WO2023217130A1 - Formulation biologique contenant des cellules de type précurseur de muscle squelettique, son procédé de préparation et son application - Google Patents
Formulation biologique contenant des cellules de type précurseur de muscle squelettique, son procédé de préparation et son application Download PDFInfo
- Publication number
- WO2023217130A1 WO2023217130A1 PCT/CN2023/092965 CN2023092965W WO2023217130A1 WO 2023217130 A1 WO2023217130 A1 WO 2023217130A1 CN 2023092965 W CN2023092965 W CN 2023092965W WO 2023217130 A1 WO2023217130 A1 WO 2023217130A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cells
- skeletal muscle
- muscle precursor
- precursor
- preparation containing
- Prior art date
Links
- 210000002027 skeletal muscle Anatomy 0.000 title claims abstract description 203
- 238000002360 preparation method Methods 0.000 title claims abstract description 51
- 239000000203 mixture Substances 0.000 title abstract description 5
- 238000009472 formulation Methods 0.000 title abstract 2
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 claims abstract description 14
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 claims abstract description 14
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 claims abstract description 14
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 claims abstract description 14
- 210000004027 cell Anatomy 0.000 claims description 205
- 239000002609 medium Substances 0.000 claims description 46
- 230000008672 reprogramming Effects 0.000 claims description 41
- 238000000034 method Methods 0.000 claims description 22
- 102000004142 Trypsin Human genes 0.000 claims description 19
- 108090000631 Trypsin Proteins 0.000 claims description 19
- 230000029087 digestion Effects 0.000 claims description 19
- 239000012588 trypsin Substances 0.000 claims description 19
- 208000029549 Muscle injury Diseases 0.000 claims description 17
- 235000015872 dietary supplement Nutrition 0.000 claims description 16
- 101000935043 Homo sapiens Integrin beta-1 Proteins 0.000 claims description 13
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 claims description 13
- 101000601661 Homo sapiens Paired box protein Pax-7 Proteins 0.000 claims description 13
- 102100025304 Integrin beta-1 Human genes 0.000 claims description 13
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 claims description 13
- 102100037503 Paired box protein Pax-7 Human genes 0.000 claims description 13
- 101000884271 Homo sapiens Signal transducer CD24 Proteins 0.000 claims description 12
- 102100038081 Signal transducer CD24 Human genes 0.000 claims description 12
- 102000004887 Transforming Growth Factor beta Human genes 0.000 claims description 11
- 108090001012 Transforming Growth Factor beta Proteins 0.000 claims description 11
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 11
- 239000000556 agonist Substances 0.000 claims description 11
- 239000003102 growth factor Substances 0.000 claims description 11
- 239000003112 inhibitor Substances 0.000 claims description 11
- 229940043355 kinase inhibitor Drugs 0.000 claims description 11
- 239000003757 phosphotransferase inhibitor Substances 0.000 claims description 11
- 239000011435 rock Substances 0.000 claims description 11
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical group C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 claims description 11
- 102000003390 tumor necrosis factor Human genes 0.000 claims description 11
- 230000004156 Wnt signaling pathway Effects 0.000 claims description 10
- 102000015696 Interleukins Human genes 0.000 claims description 9
- 108010063738 Interleukins Proteins 0.000 claims description 9
- 230000011664 signaling Effects 0.000 claims description 9
- 238000010171 animal model Methods 0.000 claims description 8
- 230000004927 fusion Effects 0.000 claims description 7
- 230000032459 dedifferentiation Effects 0.000 claims description 6
- 210000002363 skeletal muscle cell Anatomy 0.000 claims description 6
- 239000007640 basal medium Substances 0.000 claims description 5
- 239000003124 biologic agent Substances 0.000 claims description 5
- 239000003937 drug carrier Substances 0.000 claims description 5
- 238000001727 in vivo Methods 0.000 claims description 5
- 101710112752 Cytotoxin Proteins 0.000 claims description 3
- 231100000599 cytotoxic agent Toxicity 0.000 claims description 3
- 239000002619 cytotoxin Substances 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 206010028289 Muscle atrophy Diseases 0.000 abstract description 13
- 230000020763 muscle atrophy Effects 0.000 abstract description 9
- 201000000585 muscular atrophy Diseases 0.000 abstract description 9
- 230000002068 genetic effect Effects 0.000 abstract description 2
- 210000003205 muscle Anatomy 0.000 description 36
- 238000010586 diagram Methods 0.000 description 25
- 239000003550 marker Substances 0.000 description 22
- 241000699670 Mus sp. Species 0.000 description 21
- 230000014509 gene expression Effects 0.000 description 19
- 239000000243 solution Substances 0.000 description 19
- 210000001665 muscle stem cell Anatomy 0.000 description 16
- 210000001087 myotubule Anatomy 0.000 description 16
- 239000000872 buffer Substances 0.000 description 12
- 210000000651 myofibroblast Anatomy 0.000 description 11
- 210000001519 tissue Anatomy 0.000 description 11
- 229940079593 drug Drugs 0.000 description 10
- 239000003814 drug Substances 0.000 description 10
- 102000001039 Dystrophin Human genes 0.000 description 9
- 108010069091 Dystrophin Proteins 0.000 description 9
- 239000007924 injection Substances 0.000 description 9
- 238000002347 injection Methods 0.000 description 9
- 241000702421 Dependoparvovirus Species 0.000 description 8
- 206010013801 Duchenne Muscular Dystrophy Diseases 0.000 description 8
- 230000006378 damage Effects 0.000 description 8
- 229940090044 injection Drugs 0.000 description 8
- 230000001575 pathological effect Effects 0.000 description 8
- 102000004889 Interleukin-6 Human genes 0.000 description 7
- 108090001005 Interleukin-6 Proteins 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 238000000338 in vitro Methods 0.000 description 7
- 238000002560 therapeutic procedure Methods 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 6
- 239000012091 fetal bovine serum Substances 0.000 description 6
- 229940100601 interleukin-6 Drugs 0.000 description 6
- 210000000663 muscle cell Anatomy 0.000 description 6
- 230000035772 mutation Effects 0.000 description 6
- 210000003098 myoblast Anatomy 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 208000002267 Anti-neutrophil cytoplasmic antibody-associated vasculitis Diseases 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 108090000630 Oncostatin M Proteins 0.000 description 5
- 208000010886 Peripheral nerve injury Diseases 0.000 description 5
- 102100040247 Tumor necrosis factor Human genes 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 5
- 230000004220 muscle function Effects 0.000 description 5
- 210000003365 myofibril Anatomy 0.000 description 5
- 238000010186 staining Methods 0.000 description 5
- 238000002054 transplantation Methods 0.000 description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 102000009024 Epidermal Growth Factor Human genes 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- 208000029578 Muscle disease Diseases 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 238000002659 cell therapy Methods 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 210000002808 connective tissue Anatomy 0.000 description 4
- 101150015424 dmd gene Proteins 0.000 description 4
- 238000010362 genome editing Methods 0.000 description 4
- 210000002865 immune cell Anatomy 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 230000004054 inflammatory process Effects 0.000 description 4
- 230000007774 longterm Effects 0.000 description 4
- 238000010172 mouse model Methods 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000008439 repair process Effects 0.000 description 4
- 238000001356 surgical procedure Methods 0.000 description 4
- LOGFVTREOLYCPF-KXNHARMFSA-N (2s,3r)-2-[[(2r)-1-[(2s)-2,6-diaminohexanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxybutanoic acid Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]1CCCN1C(=O)[C@@H](N)CCCCN LOGFVTREOLYCPF-KXNHARMFSA-N 0.000 description 3
- AQGNHMOJWBZFQQ-UHFFFAOYSA-N CT 99021 Chemical compound CC1=CNC(C=2C(=NC(NCCNC=3N=CC(=CC=3)C#N)=NC=2)C=2C(=CC(Cl)=CC=2)Cl)=N1 AQGNHMOJWBZFQQ-UHFFFAOYSA-N 0.000 description 3
- 102000029816 Collagenase Human genes 0.000 description 3
- 108060005980 Collagenase Proteins 0.000 description 3
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 3
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 3
- 206010016654 Fibrosis Diseases 0.000 description 3
- 102000003777 Interleukin-1 beta Human genes 0.000 description 3
- 108090000193 Interleukin-1 beta Proteins 0.000 description 3
- 206010048654 Muscle fibrosis Diseases 0.000 description 3
- 206010028851 Necrosis Diseases 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 230000002159 abnormal effect Effects 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 210000002469 basement membrane Anatomy 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 210000004271 bone marrow stromal cell Anatomy 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 3
- 229960002327 chloral hydrate Drugs 0.000 description 3
- 229960002424 collagenase Drugs 0.000 description 3
- 230000008602 contraction Effects 0.000 description 3
- 229960004397 cyclophosphamide Drugs 0.000 description 3
- 229940108605 cyclophosphamide injection Drugs 0.000 description 3
- 230000007547 defect Effects 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 230000004761 fibrosis Effects 0.000 description 3
- 238000001415 gene therapy Methods 0.000 description 3
- 230000028993 immune response Effects 0.000 description 3
- 230000033001 locomotion Effects 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 3
- 230000017074 necrotic cell death Effects 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 210000004940 nucleus Anatomy 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 210000002235 sarcomere Anatomy 0.000 description 3
- 210000002435 tendon Anatomy 0.000 description 3
- 206010003694 Atrophy Diseases 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
- 101800003838 Epidermal growth factor Proteins 0.000 description 2
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 2
- 206010064571 Gene mutation Diseases 0.000 description 2
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 2
- 229930182566 Gentamicin Natural products 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 230000037444 atrophy Effects 0.000 description 2
- 210000003855 cell nucleus Anatomy 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 230000007850 degeneration Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 229940116977 epidermal growth factor Drugs 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 230000008014 freezing Effects 0.000 description 2
- 238000007710 freezing Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 229960002518 gentamicin Drugs 0.000 description 2
- 239000003862 glucocorticoid Substances 0.000 description 2
- 230000002458 infectious effect Effects 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 210000004379 membrane Anatomy 0.000 description 2
- 210000005036 nerve Anatomy 0.000 description 2
- 238000010899 nucleation Methods 0.000 description 2
- 230000000399 orthopedic effect Effects 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000011476 stem cell transplantation Methods 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 229940037128 systemic glucocorticoids Drugs 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 102000004420 Creatine Kinase Human genes 0.000 description 1
- 108010042126 Creatine kinase Proteins 0.000 description 1
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 1
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 1
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 description 1
- 102000006354 HLA-DR Antigens Human genes 0.000 description 1
- 108010058597 HLA-DR Antigens Proteins 0.000 description 1
- -1 HLA-DRPQ Proteins 0.000 description 1
- 102100023915 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 102000004310 Ion Channels Human genes 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 108020004485 Nonsense Codon Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 102000004140 Oncostatin M Human genes 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 208000026214 Skeletal muscle atrophy Diseases 0.000 description 1
- 102000013814 Wnt Human genes 0.000 description 1
- 108050003627 Wnt Proteins 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 210000004504 adult stem cell Anatomy 0.000 description 1
- 238000011166 aliquoting Methods 0.000 description 1
- 230000000735 allogeneic effect Effects 0.000 description 1
- 229940126574 aminoglycoside antibiotic Drugs 0.000 description 1
- 239000002647 aminoglycoside antibiotic agent Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000002300 anti-fibrosis Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 239000003131 biological toxin Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 230000009460 calcium influx Effects 0.000 description 1
- 230000002612 cardiopulmonary effect Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 229940126864 fibroblast growth factor Drugs 0.000 description 1
- 230000037433 frameshift Effects 0.000 description 1
- 238000003209 gene knockout Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 238000001794 hormone therapy Methods 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 238000011133 mesenchymal stem cell therapy Methods 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 238000001964 muscle biopsy Methods 0.000 description 1
- 230000009756 muscle regeneration Effects 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- 210000000107 myocyte Anatomy 0.000 description 1
- 208000018360 neuromuscular disease Diseases 0.000 description 1
- 230000037434 nonsense mutation Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 208000030212 nutrition disease Diseases 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 210000000578 peripheral nerve Anatomy 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004202 respiratory function Effects 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 210000003705 ribosome Anatomy 0.000 description 1
- 210000000518 sarcolemma Anatomy 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 230000025185 skeletal muscle atrophy Effects 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000009168 stem cell therapy Methods 0.000 description 1
- 238000009580 stem-cell therapy Methods 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 210000002303 tibia Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/067—Hepatocytes
- C12N5/0672—Stem cells; Progenitor cells; Precursor cells; Oval cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/22—Urine; Urinary tract, e.g. kidney or bladder; Intraglomerular mesangial cells; Renal mesenchymal cells; Adrenal gland
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/34—Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/36—Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/37—Digestive system
- A61K35/38—Stomach; Intestine; Goblet cells; Oral mucosa; Saliva
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/37—Digestive system
- A61K35/407—Liver; Hepatocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/37—Digestive system
- A61K35/413—Gall bladder; Bile
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/42—Respiratory system, e.g. lungs, bronchi or lung cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/52—Sperm; Prostate; Seminal fluid; Leydig cells of testes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/08—Drugs for disorders of the urinary system of the prostate
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P21/00—Drugs for disorders of the muscular or neuromuscular system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/15—Transforming growth factor beta (TGF-β)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/40—Regulators of development
- C12N2501/415—Wnt; Frizzeled
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/70—Enzymes
- C12N2501/72—Transferases (EC 2.)
- C12N2501/727—Kinases (EC 2.7.)
Definitions
- the present invention relates to the field of biotechnology, and in particular to a biological agent containing skeletal muscle precursor-like cells and its preparation method and application.
- Skeletal muscle is the largest organ of the human body, and its main functions are movement, shape maintenance, and respiration.
- myofiber cell the English name of myofiber cell is: myofiber. It is composed of many parallel-arranged myofibrils (myofibril's English name: myofibril) and is surrounded by a myofibrillar membrane (myofibril's English name: sarcolemma, the cell membrane of myofiber cells).
- myofibrillar membrane myofibril's English name: sarcolemma, the cell membrane of myofiber cells.
- basement membrane the English name of basement membrane is: basement membrane).
- each myofibril can be divided into multiple sarcomeres (the English name of sarcomeres is: sarcomeres), which generate tensile force through contraction.
- each muscle fiber cell usually contains hundreds of cell nuclei (the English name of the cell nucleus is: myonucleus). They are distributed close to the muscle fiber membrane and jointly control the movement and metabolic processes of the muscle fiber cells.
- myofiber cells skeletal muscle also contains a large amount of connective tissue (the English name of connective tissue is: connective tissue), Blood vessels, nerves, and connective tissue connect muscle fiber cells into an overall functional unit and are fixed to the bones through tendons (the English name of tendons is: tendon).
- muscle stem cells can be labeled into two groups.
- the first group of muscle stem cells accounts for about 80% of the total number. They proliferate, differentiate, fuse and eventually form myofiber cells; the scientists will The second group of muscle stem cells is called "reserve cells”.
- the second group of muscle stem cells divides at a significantly slower rate and eventually becomes a muscle stem cell bank in muscle tissue.
- follow-up studies have shown that the first group of muscle stem cells and the second group of muscle stem cells come from the asymmetric division of muscle stem cells (the English name of asymmetric division is: asymmetric division).
- Duchenne muscular dystrophy is a common fatal neuromuscular disease in children. It is inherited in an X-linked recessive manner and is caused by mutations in the DMD gene. Children often develop the disease in childhood. The disease is hidden and can easily be missed or misdiagnosed in the early stages. Because defects at the genetic level are irreversible, the body cannot repair the damage through muscle cell regeneration and compensation, which gradually leads to late changes in massive fibrosis in skeletal muscles, myocardium and other parts of the body, and ultimately leads to death due to severe impact on the cardiopulmonary function of the child.
- the DMD gene is located at Xp21, with a genome span of 2.4Mb.
- Dystrophin deficiency leads to at least two results. The first result is that muscle cells are destroyed by mechanical force during eccentric contraction, and the ion channels that are sensitive to mechanical force are dysregulated. The second result is that muscle cells are damaged by mechanical force during eccentric contraction. Because the muscle cell membrane cannot remain intact, abnormal protein and calcium influx, creatine kinase efflux, etc. occur, eventually causing muscle cell degeneration and necrosis.
- Peripheral nerve injury (English name: peripheral nerve injury, PNI) is a common clinical trauma, which can be caused by various reasons such as tearing, traction, cutting, compression, ischemia, high temperature, freezing, infection and nutritional and metabolic disorders. Peripheral nerves can regenerate to a certain extent after damage, but the recovery rate is very slow. After nerves are severely damaged, the skeletal muscles they control atrophy within a few days. As the atrophy time prolongs, cell apoptosis gradually increases, and the number of myocyte nuclei gradually decreases, eventually causing permanent loss of skeletal muscle function.
- PNI peripheral nerve injury
- Muscle atrophy caused by genetic mutations such as Duchenne muscular dystrophy and muscle atrophic changes caused by peripheral nerve injury (PNI) will cause muscle atrophic lesions, causing the number of muscle cells to decrease until the permanent loss of skeletal muscle function thereby threatening the patient's life. If muscle atrophy can be controlled, muscle degeneration can be reduced and movement and respiratory functions can be saved. Therefore, there are several ideas for the treatment of muscle diseases: cell therapy, gene editing therapy and traditional hormone therapy.
- MTT myoblast transfer therapy
- the myofiber cells fuse and then express normal DMD, correcting the degeneration, necrosis and other changes of muscle fibers; mesenchymal stem cells (English name: mesenchymal stem cells, English abbreviation: MSC) are a type of adult stem cells with multiple differentiation potentials , MSCs cultured in vitro for more than 3 generations have good uniformity, low immune response to vein transplantation, and 5 When MSCs were transferred to gene knockout mice, the intensity of DMD immunofluorescence increased with time, confirming the feasibility of introducing and expressing the DMD gene through this approach.
- mesenchymal stem cells English name: mesenchymal stem cells, English abbreviation: MSC
- the biggest difficulty in muscle stem cell therapy is the lack of donor cells.
- the patient's own muscle stem cells need to be removed, and the expression of Dystrophin can be obtained through infection in vitro.
- Injection back into the patient's body but there is currently no method that can expand large amounts of human muscle stem cells in vitro;
- research on combined mesenchymal stem cell therapy shows that this method is only suitable for patients with mild to moderate muscle atrophy, but not for patients with extreme muscle mass. The curative effect is minimal, and the long-term efficacy is unknown.
- Gene editing therapy In vivo gene editing mediated by adeno-associated virus (English abbreviation: AAV) is a promising treatment idea. By injecting adeno-associated virus, the expression of dystrophin can be restored in myofibroblasts to a certain extent, thus To increase the strength of muscle fiber cells and avoid necrosis, the advantages of AAV treatment are as follows: first, the virus can be amplified in vitro and is easy to obtain; second, AAV has been proven to have low tumorigenicity in vivo and is relatively safe; third, because the virus Due to its diffusibility, AAV can infect a larger range of tissues through blood transport; other gene editing therapies also include exon skipping and stop codon readthrough.
- Exon skipping uses exon skipping splicing technology to frameshift DMD. Changing the mutation type to an integer mutation type may transform DMD with severe symptoms into mild symptoms; terminate DMD Code reading: The aminoglycoside antibiotic gentamicin can reduce the ability of ribosomes to recognize abnormal terminators during the translation process of dystrophin, and skip these abnormal terminators to continue translation. This result was found in mouse animal experiments It also achieved significant curative effect.
- Gene therapy has the following shortcomings: First, the biggest obstacle to adeno-associated virus therapy is that it cannot break through the microenvironmental barrier to infect muscle stem cells. Myofibroblasts that have been infected by the virus and acquire Dystrophin expression will die due to lifespan and metabolic replacement, so AAV therapy cannot fundamentally solve the problem of muscle diseases and can only treat patients multiple times and continuously; secondly, exon skipping can only target specific gene defect types, and the energy efficiency of oligonucleotides entering the cell membrane is low, which may It is immunogenic; thirdly, stop codon read-through therapy only targets nonsense mutations because gentamicin has low read-through energy efficiency and long-term large-scale application has large toxic side effects, and the drug may have short-term or long-term side effects.
- Glucocorticoids have achieved certain success in the treatment of DMD due to their anti-inflammatory effects and are most commonly used in clinical applications. However, the side effects of glucocorticoids often outweigh their benefits and can produce long-term inflammatory reactions that require combined anti-inflammatory treatment.
- the purpose of the present invention is to provide a biological preparation containing skeletal muscle precursor-like cells and its preparation method and application, so as to solve the problem of immune rejection in the treatment of muscle atrophy caused by gene mutations in the prior art.
- the present invention provides a biological preparation containing skeletal muscle precursor-like cells that negatively express at least one of CD34, CD45, and HLA-DRPQ.
- the beneficial effect of the biological preparation containing skeletal muscle precursor-like cells of the present invention is that the negative expression of at least one of CD34, CD45, and HLA-DRPQ by the skeletal muscle precursor-like cells allows the human body to respond to the skeletal muscle.
- Precursor-like cells are well accepted, so this application solves the problem of immune rejection in the existing technology for treating muscle atrophy caused by gene mutations.
- the skeletal muscle precursor-like cells positively express at least one of CD29, CD56, CD24 and PAX7.
- the present invention also provides a method for preparing a biological preparation containing skeletal muscle precursor-like cells, which includes the following steps:
- S0 Provides skeletal muscle precursor-like cells
- S1 Mix the skeletal muscle precursor-like cells with a pharmaceutically acceptable carrier to obtain a biological preparation containing skeletal muscle precursor-like cells, wherein the skeletal muscle precursor-like cells positively express CD29, CD56, At least one of CD24 and PAX7, and the skeletal muscle precursor-like cells negatively express at least one of CD34, CD45, and HLA-DRPQ.
- the beneficial effect of the preparation method of the biological preparation containing skeletal muscle precursor-like cells of the present invention is that the preparation method of the biological preparation containing skeletal muscle precursor-like cells is simple, and the obtained biological preparation containing skeletal muscle precursor-like cells It can reduce the impact of muscle damage on the human body, adapt well to the human body, and will not cause immune rejection with the recipient.
- the preparation method of skeletal muscle precursor-like cells includes the following steps:
- S01 Place the primary skeletal muscle cells into a reprogramming medium for dedifferentiation culture until the fusion degree of the skeletal muscle precursor-like cells is not less than 80%, and use trypsin digestion solution to treat the skeletal muscle
- the precursor-like cells are digested to obtain skeletal muscle precursor-like cells, wherein the reprogramming medium includes basal medium, tumor necrosis factor, interleukin and oncostatin.
- the preparation method of the skeletal muscle precursor-like cells also includes: S02: placing the skeletal muscle precursor-like cells into the reprogramming medium for expansion and culture until the skeletal muscle precursor-like cells After the fusion degree of the cells is not less than 80%, the skeletal muscle precursor-like cells are digested using the trypsin digestion solution to obtain passaged skeletal muscle precursor-like cells.
- S02 placing the skeletal muscle precursor-like cells into the reprogramming medium for expansion and culture until the skeletal muscle precursor-like cells After the fusion degree of the cells is not less than 80%, the skeletal muscle precursor-like cells are digested using the trypsin digestion solution to obtain passaged skeletal muscle precursor-like cells.
- the beneficial effect is that skeletal muscle precursor-like cells can be continuously expanded in the reprogramming medium and the obtained skeletal muscle precursor-like cells can express skeletal muscle precursor-like cell markers.
- the reprogramming medium also includes growth factors, ROCK kinase inhibitors, Wnt signaling pathway agonists, TGF- ⁇ signaling inhibitors and nutritional supplements.
- the content of tumor necrosis factor is 2-20ng/mL
- the content of interleukin is 20-50ng/mL
- the content of oncostatin is based on the volume of the reprogramming medium. is 1-10ng/mL.
- the content of the growth factor is 40-80ng/mL
- the content of the ROCK kinase inhibitor is 5-20 ⁇ M
- the content of the Wnt signaling pathway agonist is is 1-10 ⁇ M
- the content of the TGF- ⁇ signal inhibitor is 0.1-5 ⁇ M
- the content of the nutritional supplement is 0.5-10% of the final volume of the reprogramming medium.
- the present invention also provides an application of a biological preparation containing skeletal muscle precursor-like cells, using the biological preparation containing skeletal muscle precursor-like cells to intervene in an in vivo animal model.
- the beneficial effects of the application of the skeletal muscle precursor-like cells of the present invention are: after infusion of biological agents containing skeletal muscle precursor-like cells, local inflammation can be well controlled, muscle fibrosis can be controlled, and muscle degeneration can be alleviated. sexual lesions so that skeletal muscles can recover better.
- the animal model includes a cytotoxin-induced muscle injury model.
- Figure 1 is a photographic schematic diagram of the morphology of the first generation skeletal muscle precursor-like cells according to an embodiment of the present invention
- Figure 2 is a photographic schematic diagram of the morphology of fourth-generation skeletal muscle precursor-like cells according to an embodiment of the present invention
- Figure 3 is a schematic diagram of the proliferation curve of skeletal muscle precursor-like cells according to an embodiment of the present invention.
- Figure 4 is a schematic diagram of the gene expression marker CD34 of skeletal muscle precursor-like cells according to an embodiment of the present invention.
- Figure 5 is a schematic diagram of the gene expression marker CD45 of skeletal muscle precursor-like cells according to an embodiment of the present invention.
- Figure 6 shows gene expression markers of skeletal muscle precursor-like cells according to the embodiment of the present invention. Schematic diagram of HLA-DRPQ situation
- Figure 7 is a schematic diagram of the gene expression marker CD29 of skeletal muscle precursor-like cells according to an embodiment of the present invention.
- Figure 8 is a schematic diagram of the gene expression marker CD56 of skeletal muscle precursor-like cells according to an embodiment of the present invention.
- Figure 9 is a schematic diagram of the gene expression marker CD24 of skeletal muscle precursor-like cells according to an embodiment of the present invention.
- Figure 10 is a schematic diagram of the gene expression marker PAX7 in skeletal muscle precursor-like cells according to an embodiment of the present invention.
- Figure 11 is a schematic diagram comparing photos of pathological tissue sections of the drug-induced muscle injury model mice in the experimental group and the control group at different times according to the embodiment of the present invention
- Figure 12 is a comparative diagram of the area distribution of new myofibroblasts in the experimental group and the control group of drug-induced muscle injury model mice according to the embodiment of the present invention.
- Fibrosis is a significant pathological feature seen in muscle biopsy in patients with muscle atrophy, which can lead to muscle function disorders and ultimately fatality; anti-fibrosis treatment can not only improve muscle function, but also promote muscle regeneration, gene introduction and stem cells The colonization efficiency will become a necessary supplement for future gene and cell therapy.
- anti-fibrosis treatment can not only improve muscle function, but also promote muscle regeneration, gene introduction and stem cells The colonization efficiency will become a necessary supplement for future gene and cell therapy.
- drugs targeting fibrosis were tested and shown to improve muscle function and muscle damage phenotypes.
- the biological agent containing skeletal muscle precursor-like cells of the present invention after being reinfused into the body, can effectively control local inflammation, control muscle fibrosis, and reduce muscle degeneration, so that skeletal muscles can be better treated. recover.
- the present invention provides a biological preparation comprising skeletal muscle precursor-like cells that negatively express at least one of CD34, CD45, and HLA-DRPQ.
- the skeletal muscle precursor-like cells negatively express at least one of CD34, CD45, and HLA-DRPQ, so that the human body can accept the skeletal muscle precursor-like cells well, so that this application solves the problem of the existing technology.
- the HLA-DRPQ is the abbreviation of HLA-DR/DP/DQ.
- the skeletal muscle precursor-like cells positively express at least one of CD29, CD56, CD24 and PAX7.
- the present invention also provides a method for preparing a biological preparation containing skeletal muscle precursor-like cells, which includes the following steps:
- S0 Provides skeletal muscle precursor-like cells
- S1 Mix the skeletal muscle precursor-like cells with a pharmaceutically acceptable carrier to obtain a biological preparation containing skeletal muscle precursor-like cells, wherein the skeletal muscle precursor-like cells positively express CD29, CD56, At least one of CD24 and PAX7, and the skeletal muscle precursor-like cells negatively express at least one of CD34, CD45, and HLA-DRPQ.
- the preparation method of the biological preparation containing skeletal muscle precursor-like cells is simple, and the obtained biological preparation containing skeletal muscle precursor-like cells can reduce the impact of muscle damage on the human body, and can adapt well to the human body without affecting the human body. Can cause immune rejection in the recipient.
- the pharmaceutically acceptable carrier includes but is not limited to physiological saline and compound electrolyte injection.
- the method for preparing skeletal muscle precursor-like cells includes the following steps:
- S01 Place the primary skeletal muscle cells into a reprogramming medium for dedifferentiation culture until the fusion degree of the skeletal muscle precursor-like cells is not less than 80%, and use trypsin digestion solution to treat the skeletal muscle
- the precursor-like cells are digested to obtain skeletal muscle precursor-like cells, wherein the reprogramming medium includes basal medium, tumor necrosis factor, interleukin and oncostatin.
- the skeletal muscle primary cells are put into a reprogramming medium for dedifferentiation culture until the fusion degree of the skeletal muscle precursor-like cells is not less than 80%, and the trypsin digestion solution is used to digest the cells.
- Skeletal muscle precursor-like cells are digested to obtain skeletal muscle precursors -like cells, wherein the reprogramming medium includes basal medium, tumor necrosis factor, interleukin and oncostatin, so that the skeletal muscle precursor-like cells continuously achieve self-renewal and proliferation, thereby solving the existing problem
- the problem is that culture technology cannot culture skeletal muscle precursor-like cells in large quantities in vitro.
- the preparation method of the skeletal muscle precursor-like cells further includes: S02: Put the skeletal muscle precursor-like cells into the reprogramming medium for expansion and culture until the skeletal muscle precursor-like cells are After the fusion degree of the muscle precursor-like cells is not less than 80%, the skeletal muscle precursor-like cells are digested using the trypsin digestion solution to obtain passaged skeletal muscle precursor-like cells. Skeletal muscle precursor-like cells can be continuously expanded in the reprogramming medium and the resulting skeletal muscle precursor-like cells can express skeletal muscle precursor-like cell markers.
- the reprogramming medium further includes growth factors, ROCK kinase inhibitors, Wnt signaling pathway agonists, TGF- ⁇ signaling inhibitors and nutritional supplements.
- the content of the tumor necrosis factor is 2-20ng/mL
- the content of the interleukin is 20-50ng/mL
- the inhibitory factor is 20-50ng/mL.
- the content of oncostin is 1-10ng/mL.
- the content of the tumor necrosis factor is 5-15ng/mL
- the content of the interleukin is 25-40ng/mL
- the inhibitory factor is 25-40ng/mL.
- the content of oncostin is 2-8ng/mL.
- the English name of the tumor necrosis factor is tumor necrosis factor
- the English abbreviation of the tumor necrosis factor is TNF
- the tumor necrosis factor TNF originates from the coastal area of Shanghai.
- the interleukin includes interleukin-1 ⁇ and interleukin-6.
- the English abbreviation of interleukin-1 ⁇ is IL-1 ⁇ .
- the source of interleukin-1 ⁇ is From Huamei Biotechnology; the English abbreviation of interleukin-6 is IL-6, and the interleukin-6 is derived from Huamei Biotechnology.
- the English name of the oncostatin is oncostatin M
- the English abbreviation of the oncostatin is OSM
- the oncostatin is derived from Yisheng Biotechnology.
- the content of the growth factor is 40-80ng/mL
- the content of the ROCK kinase inhibitor is 5-20 ⁇ M
- the Wnt signaling pathway The content of the agonist is 1-10 ⁇ M
- the content of the TGF- ⁇ signaling inhibitor is 0.1-5 ⁇ M
- the content of the nutritional supplement is 0.5-10% of the final volume of the reprogramming medium.
- the content of the growth factor is 60-75ng/mL
- the content of the ROCK kinase inhibitor is 5-15 ⁇ M
- the Wnt signal The content of the pathway agonist is 2-6 ⁇ M
- the content of the TGF- ⁇ signaling inhibitor is 1-3 ⁇ M
- the content of the nutritional supplement is 1-7% of the final volume of the reprogramming medium.
- the reprogramming medium further includes growth factors, ROCK kinase inhibitors, Wnt signaling pathway agonists, TGF- ⁇ signaling inhibitors, nutritional supplements and buffers.
- the growth factors include epidermal growth factor and alkaline Fibroblast growth factor, the epidermal growth factor is derived from nearshore organisms, and the basic fibroblast growth factor is derived from nearshore organisms; the ROCK kinase inhibitor Y-27632 is derived from Taoshu organisms, and the Wnt signaling pathway
- the agonist CHIR99021 is derived from Taoshu Biotech
- the TGF- ⁇ signaling inhibitor A8301 is derived from Taoshu Biotech
- the nutritional supplements include N2 nutritional supplements and B27 nutritional supplements, the N2 nutritional supplements are derived from Yisheng biology.
- the present invention also provides an application of a biological preparation containing skeletal muscle precursor-like cells, using the biological preparation containing skeletal muscle precursor-like cells to intervene in an in vivo animal model.
- skeletal muscle precursor-like cells Through the infusion of biological agents containing skeletal muscle precursor-like cells, local inflammation can be well controlled, muscle fibrosis can be controlled, and muscle degeneration can be alleviated, so that skeletal muscles can be better recovered.
- the animal model includes a cytotoxin-induced muscle injury model.
- Human skeletal muscle precursor-like cells that positively express CD29, CD56 and PAX7 are obtained from skeletal muscle tissue removed after clinical orthopedic surgery or donated by donors.
- the skeletal muscle tissue was found to be normal skeletal muscle tissue upon pathological examination.
- the skeletal muscle tissue removed after clinical orthopedic surgery or the skeletal muscle tissue donated by a donor is a surgical sample derived from a patient aged no more than 40 years old.
- the patient has no infectious virus infection after medical examination, and the patient has no infectious virus infection before surgery. No steroid use within 6 months drug.
- the patient was fully informed about the purpose of obtaining surgical samples before surgery and signed an informed consent form.
- sterile PBS buffer (0.01M) to clean and sterilize the skeletal muscle tissue
- cell digestion solution 3 ml of cell digestion solution to digest the skeletal muscle tissue at 37 degrees Celsius for 90 minutes to obtain primary skeletal muscle cells.
- the cell digestion solution is composed of type III collagenase, sterile PBS buffer and trypsin digestion solution, and based on the volume of the cell digestion solution, it includes 49.5% sterile PBS buffer (0.01M) and Trypsin digestion solution (1X) with a content of 49.5%, type III collagenase with a content of 1%, type III collagenase is from Shanghai Maokang Biotechnology Co., Ltd., and sterile PBS buffer is from Wuhan Pronosai Life Technology Co., Ltd., trypsin digestion liquid (containing 0.25% trypsin) comes from Biyuntian;
- the skeletal muscle primary cell suspension was screen sorted with the assistance of sterile PBS buffer using a 70-micron sterile mesh to remove mucus and undigested tissue and collect the filtrate to complete the screen sorting ;
- the filtrate is centrifuged and the supernatant is removed to obtain a precipitate.
- Red blood cell lysate (from Soleba) is added to the precipitate for resuspension. The process is centrifuged again and the above process is repeated until the precipitate is centrifuged again. Until no red blood cells are observed in the sediment, the red blood cells are lysed and removed to finally obtain primary skeletal muscle cells.
- the centrifugation speed for each centrifugation process is 1000g, and the centrifugation time for each centrifugation process is 3 minutes.
- the composition of the reprogramming medium includes: based on the volume of the reprogramming medium, basic culture Base Ham F10 (from Wuhan Punosai Life Technology Co., Ltd.) accounts for 91% of the total volume of the reprogramming medium, with a content of 20 ng/ml of epithelial cell growth factor EGF and a content of 50 ng/ml of alkaline Fibroblast Growth Factor bFGF, 1% Nutritional Supplement N2 (1X), 1% Nutritional Supplement B27 (1X), 10 ⁇ M ROCK Kinase Inhibitor Y-27632, 3 ⁇ M Wnt Signaling pathway agonist CHIR99021, TGF- ⁇ signaling inhibitor A8301 at 1 ⁇ M, Tumor necrosis factor TNF at 10 ng/ml, Interleukin-1b at 20 ng/ml, 5 ng /ml of oncostatin OSM, 10 ng/ml of interleukin-6, 5% fetal bovine serum (the English abbrevi
- Control medium Based on the volume of the control medium, the basal medium Ham F10 (from Wuhan Prosai Life Technology Co., Ltd.) accounts for 91% of the total volume of the control medium, with a content of 20 ng/ml of epithelial cells.
- EGF EGF
- basic fibroblast growth factor bFGF at 50 ng/ml
- 1% nutritional supplement N2 (1X) 1% nutritional supplement B27 (1X) at 10 ⁇ M ROCK kinase inhibitor Y-27632, Wnt signaling pathway agonist CHIR99021 at 3 ⁇ M, TGF- ⁇ signaling inhibitor A8301 at 1 ⁇ M
- 5% fetal bovine serum the English abbreviation of fetal bovine serum is: FBS , from Beijing Baiolaibo Technology Co., Ltd.).
- the difference between reprogramming medium and control medium contains tumor necrosis factor TNF at 10 ng/ml, interleukin-1b at 20 ng/ml, and 5 ng/ml.
- ml of oncostatin OSM containing 10 ng/ml of interleukin-6, while the control medium did not contain tumor necrosis factor TNF, interleukin-1b, Oncostatin OSM and interleukin-6.
- Figure 1 is a schematic photographic diagram of the morphology of the first-generation skeletal muscle precursor-like cells according to an embodiment of the present invention
- Figure 2 is a schematic photographic diagram of the morphology of the fourth-generation skeletal muscle precursor-like cells according to an embodiment of the present invention
- Figure 3 is a schematic diagram of the implementation of the present invention. Schematic diagram of the proliferation curve of skeletal muscle precursor-like cells of an example.
- skeletal muscle primary cells Place skeletal muscle primary cells in a 6-well plate at a seeding area of 10,000 cells/cm2, add 2 ml of reprogramming medium to each well for dedifferentiation culture to obtain skeletal muscle precursor-like cells until skeletal muscle precursor-like cells are obtained. After the confluence of the cells is not less than 80%, the skeletal muscle precursor-like cells are digested using trypsin digestion solution. The digestion time is 5 minutes.
- the trypsin digestion solution (containing 0.25% trypsin) is derived from Biyuntian;
- the skeletal muscle precursor-like cells will continue to be subcultured using reprogramming medium to obtain the first generation (P1) skeletal muscle precursor-like cells, the second generation (P2) skeletal muscle precursor-like cells, and the second generation (P2) skeletal muscle precursor-like cells.
- Third generation (P3) skeletal muscle precursor-like cells skeletal muscle precursor-like cells.
- Tenth generation (P10) skeletal muscle precursor-like cells, skeletal muscle precursor-like cells can be continuously passaged in reprogramming culture medium.
- the digestion time is 5 minutes, in which the trypsin digestion solution (containing 0.25% trypsin) Sourced from Biyuntian; the control skeletal muscle precursor-like cells will continue to be subcultured in the control medium to obtain the first
- the second generation (P1) controls skeletal muscle precursor-like cells
- the second generation (P2) controls skeletal muscle precursor-like cells
- the third generation (P3) controls skeletal muscle precursor-like cells
- the fourth generation (P4) controls skeletal muscle precursor-like cells.
- Somatic-like cells, fifth passage (P5) control skeletal muscle precursor-like cells.
- the control skeletal muscle precursor-like cells can be passaged in the control medium, when they are passaged to the fifth passage, the control skeletal muscle precursor-like cells have Unable to proliferate in control medium, see Figure 3.
- Figure 1 shows the P1 generation skeletal muscle precursor-like cells
- Figure 2 shows the P4 generation skeletal muscle precursor-like cells.
- the skeletal muscle precursor-like cells remain consistent in shape and are polygonal.
- the reprogramming medium can allow the skeletal muscle precursor-like cells to continue to expand, and the skeletal muscle precursor-like cells
- the expansion ability of cells has always been enhanced; using control medium for subculture, the expansion ability of control skeletal muscle precursor-like cells increased first and then decreased with the increase in the number of passages, and in the second generation , has the strongest amplification ability.
- the abscissa in Figure 3 is the number of cell generations, the ordinate is the cell amplification multiple, and the number of generations is the number of passages.
- Figure 4 is a schematic diagram of the gene expression marker CD34 of skeletal muscle precursor-like cells according to an embodiment of the present invention
- Figure 5 is a schematic diagram of the gene expression marker CD45 of skeletal muscle precursor-like cells according to an embodiment of the present invention
- Figure 6 It is a schematic diagram of the gene expression marker HLA-DRPQ of skeletal muscle precursor-like cells according to an embodiment of the present invention
- Figure 7 is a schematic diagram of the gene expression marker CD29 of skeletal muscle precursor-like cells according to an embodiment of the present invention
- Figure 8 It is the gene expression marker CD56 of skeletal muscle precursor-like cells according to the embodiment of the present invention.
- Figure 9 is a schematic diagram of the gene expression marker CD24 of skeletal muscle precursor-like cells according to an embodiment of the present invention
- Figure 10 is a schematic diagram of the gene expression marker PAX7 of skeletal muscle precursor-like cells according to an embodiment of the present invention.
- the skeletal muscle precursor-like cells were washed with sterile PBS buffer (from Wuhan Prosai Life Technology Co., Ltd.), and then digested with trypsin digestion solution (Pancreatic Pancreas). Enzyme digestion solution (containing 0.25% trypsin) derived from Biyuntian) digested skeletal muscle precursor-like cells and then centrifuged to collect the cell pellet; add 600 microliters of staining buffer to the cell pellet to resuspend the cells.
- sterile PBS buffer from Wuhan Prosai Life Technology Co., Ltd.
- trypsin digestion solution Pancreatic Pancreas
- FIG 4 Figure 5
- the flow cytometry analysis results shown in Figure 6, Figure 7, Figure 8, Figure 9, and Figure 10 in which the staining buffer is from Thermo Fisher Scientific, and the flow cytometry antibody to be tested is from Thermo Fisher Scientific. company.
- the specific operations and analysis steps of flow cytometric detection are routine technical means for those skilled in the art and will not be described in detail here.
- the positivity rate of the positive peak of marker CD34 is 0.54%
- the positivity rate of the positive peak of marker CD45 is 0.97%
- the positivity rate of the positive peak of marker HLA-DRPQ is 0.68% , when the positive rate of the positive peak is greater than 70%, it means that the skeletal muscle precursor-like cells positively express the marker.
- the skeletal muscle precursor-like cells of the present invention negatively express CD34, CD45 and HLA-DRPQ; refer to Figure 7, Figure 8, Figure 9 and Figure 10, the positive rate of the negative peak of the marker CD29 is 99.9%, and the negative rate of the positive peak of the marker CD56 100%, the positive rate of the negative peak of the marker CD24 is 99.2%, and the positive rate of the negative peak of the marker PAX7 is 93.3%.
- the positive rate of the positive peak is greater than 70%, it means that skeletal muscle precursor-like cells positively express the marker. Therefore, the skeletal muscle precursor-like cells of the present invention positively express CD29, CD56, CD24 and PAX7.
- the abscissas and ordinates in Figure 4, Figure 5, Figure 6, Figure 7, Figure 8, Figure 9 and Figure 10 have the same meaning.
- the abscissa in Figure 4 represents the relative fluorescence intensity, and the ordinate represents the number of cells.
- the skeletal muscle precursor-like cells are mixed with a pharmaceutically acceptable carrier to prepare a biological preparation containing the skeletal muscle precursor-like cells.
- the biological preparation containing the skeletal muscle precursor-like cells is used to repair muscle damage, including skeletal muscle.
- the specific steps of the biological preparation of precursor-like cells include: mixing the skeletal muscle precursor-like cells with physiological saline to prepare a biological preparation containing skeletal muscle precursor-like cells, that is, a biological preparation containing skeletal muscle precursor-like cells. It is a skeletal muscle precursor-like cell injection, wherein each microliter of the skeletal muscle precursor-like cell injection contains 2 ⁇ 10 4 skeletal muscle precursor-like cells.
- Figure 11 is a schematic diagram comparing photos of pathological tissue sections of the drug-induced muscle injury model mice in the experimental group and the control group at different times according to the embodiment of the present invention
- Figure 12 is a schematic diagram of the drug-induced muscle injury model mice in the experiment according to the embodiment of the present invention. Comparison of the area distribution of new myofibroblasts in the group and control group.
- Animal models refer to drug-induced muscle injury models
- mice 8-week-old male mice were selected.
- cyclophosphamide dry powder was diluted to 10 ⁇ M with sterile PBS buffer to obtain cyclophosphamide injection. After aliquoting, store the solution at minus 80 degrees Celsius to avoid repeated freezing and thawing of cyclophosphamide injection; mice were intraperitoneally injected with 10% chloral hydrate. After injecting chloral hydrate, after one to two minutes, the mice were leveled.
- cyclophosphamide Lying on a clean table, use a shaver to remove the hair outside the tibialis anterior muscle of the mouse, and use an insulin injection needle to vertically inject 100 microliters of cyclophosphamide injection into the tibialis anterior muscle of the mouse in 3 injections.
- One needle was positioned at the center of the tibialis anterior muscle, and the remaining two needles were positioned at both ends of the tibialis anterior muscle to obtain a drug-induced muscle injury model.
- the English abbreviation of cyclophosphamide is CTX
- cyclophosphamide dry powder comes from Shanghai. Yuanxi Biotechnology Co., Ltd.
- sterile PBS buffer was from Wuhan Pronosai Life Technology Co., Ltd.
- chloral hydrate was from Shanghai Bevanta Biotechnology Co., Ltd.
- mice On day 0, 50 ⁇ l of skeletal muscle precursor-like cell injection was injected into the tibialis anterior muscle of mice. On day 7, 50 ⁇ l of skeletal muscle precursor-like cell injection was injected into the tibialis anterior muscle of mice again. Cell injection, and a part of the tibialis anterior muscles of the experimental mice were collected, and the tibialis anterior muscles were sampled for pathological HE staining to examine the damage to the tibialis anterior muscles; on the 15th day, the mice were sacrificed to collect the tibialis anterior muscles of the mice, and the tibialis anterior muscles were collected. Muscle samples were taken for pathological HE staining to examine the damage to the tibialis anterior muscle. Among them, HE staining is hematoxylin-eosin staining.
- Control group On day 0, 50 ⁇ l of normal saline was injected into the tibialis anterior muscle of mice. On day 7, 50 ⁇ l of normal saline was injected into the tibialis anterior muscle of mice again, and a part of the tibia of the experimental mice was collected.
- samples of the tibialis anterior muscle were sampled for pathological HE staining to examine the damage to the tibialis anterior muscle; on the 15th day, the mice were sacrificed to collect the tibialis anterior muscles of the mice, and samples of the tibialis anterior muscle were sampled for pathological HE staining to examine the damage to the tibialis anterior muscle.
- HE dye The color is hematoxylin-eosin staining.
- the tibialis anterior muscle of the mice in the experimental group recovered better.
- the visual field was filled with new myofibroblasts with nuclei in the center.
- the number of immune cells was smaller, indicating the immune response of the tibialis anterior muscle after muscle injury. It has basically ended; while in the tibialis anterior muscle of the control mice, there were a large number of immune cells, and at the same time, the number of new myofibroblasts was small and the cross-sectional area was small.
- the skeletal muscle precursor-like cell treatment method provided by the present invention can better solve muscle diseases. Because skeletal muscle precursor-like cells can proliferate and differentiate to produce new myofibroblasts in the body, and can also home in to form a muscle stem cell bank, they can theoretically provide patients with a steady supply of cells. Use reprogramming medium to culture skeletal muscle precursor-like cells so that the skeletal muscle precursor-like cells have the ability to continue to proliferate in vitro.
- mice in the experimental group recovered better.
- the visual field was filled with new myofibroblasts with nuclei in the center.
- the number of immune cells was smaller, indicating that the tibialis anterior muscle was damaged.
- the immune response after muscle injury has basically ended; while in the tibialis anterior muscle of mice in the control group, there were a large number of immune cells, and at the same time, the number of new myofibroblasts was small and the cross-sectional area was small.
- the surface of the skeletal muscle precursor-like cells obtained by the present invention does not express immune rejection reactions, In response to the related protein HLA-DRPQ, allogeneic reinfusion of skeletal muscle precursor-like cells can be achieved.
- the skeletal muscle precursor-like cells obtained by the present invention positively express CD29, CD56, CD24 and PAX7, and the skeletal muscle precursor-like cells can After being expanded in vitro to establish a cell seed bank, skeletal muscle precursor-like cells can be directly injected in situ into skeletal muscle precursor-like cell preparations when used to treat clinical skeletal muscle atrophy.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Developmental Biology & Embryology (AREA)
- Epidemiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Virology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Physiology (AREA)
- Genetics & Genomics (AREA)
- Nutrition Science (AREA)
- Wood Science & Technology (AREA)
- Urology & Nephrology (AREA)
- Gastroenterology & Hepatology (AREA)
- Pulmonology (AREA)
- General Engineering & Computer Science (AREA)
- Reproductive Health (AREA)
- Cardiology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Vascular Medicine (AREA)
- Pain & Pain Management (AREA)
- Rheumatology (AREA)
- Physical Education & Sports Medicine (AREA)
- Orthopedic Medicine & Surgery (AREA)
Abstract
L'invention concerne une formulation biologique contenant des cellules de type précurseur de muscle squelettique, et son procédé de préparation et son application. Les cellules de type précurseur de muscle squelettique expriment au moins négativement l'un parmi CD34, CD45 et HLA-DRPQ. Par conséquent, le corps humain peut bien accepter les cellules de type précurseur de muscle squelettique. La présente demande résout le problème, dans l'état de la technique, du rejet immunitaire dans le traitement de l'atrophie musculaire provoquée par une mutation génétique.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210503678.6 | 2022-05-10 | ||
CN202210503678 | 2022-05-10 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2023217130A1 true WO2023217130A1 (fr) | 2023-11-16 |
Family
ID=88621424
Family Applications (9)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2023/092959 WO2023217126A1 (fr) | 2022-05-10 | 2023-05-09 | Cellule de type précurseur épithélial rénal, son procédé de préparation, sa préparation et son utilisation |
PCT/CN2023/092963 WO2023217128A1 (fr) | 2022-05-10 | 2023-05-09 | Cellule de type précurseur épithélial de muqueuse gastrique, son procédé de préparation et son utilisation |
PCT/CN2023/092954 WO2023217121A1 (fr) | 2022-05-10 | 2023-05-09 | Formulation biologique contenant des cellules de type précurseur rénal, son procédé de préparation et son application |
PCT/CN2023/092958 WO2023217125A1 (fr) | 2022-05-10 | 2023-05-09 | Cellule de type précurseur de la prostate et préparation cellulaire, procédé de préparation associé et utilisation associée |
PCT/CN2023/092965 WO2023217130A1 (fr) | 2022-05-10 | 2023-05-09 | Formulation biologique contenant des cellules de type précurseur de muscle squelettique, son procédé de préparation et son application |
PCT/CN2023/092967 WO2023217132A1 (fr) | 2022-05-10 | 2023-05-09 | Procédé de préparation et utilisation d'une cellule de type précurseur de vésicule biliaire |
PCT/CN2023/092956 WO2023217123A1 (fr) | 2022-05-10 | 2023-05-09 | Procédé de préparation et utilisation d'une cellule de type précurseur pulmonaire |
PCT/CN2023/092969 WO2023217134A1 (fr) | 2022-05-10 | 2023-05-09 | Préparation biologique contenant une cellule de type précurseur myocardique, son procédé de préparation et application |
PCT/CN2023/092964 WO2023217129A1 (fr) | 2022-05-10 | 2023-05-09 | Cellule de type précurseur de conduit biliaire intrahépatique, préparation cellulaire, procédé de préparation et application |
Family Applications Before (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2023/092959 WO2023217126A1 (fr) | 2022-05-10 | 2023-05-09 | Cellule de type précurseur épithélial rénal, son procédé de préparation, sa préparation et son utilisation |
PCT/CN2023/092963 WO2023217128A1 (fr) | 2022-05-10 | 2023-05-09 | Cellule de type précurseur épithélial de muqueuse gastrique, son procédé de préparation et son utilisation |
PCT/CN2023/092954 WO2023217121A1 (fr) | 2022-05-10 | 2023-05-09 | Formulation biologique contenant des cellules de type précurseur rénal, son procédé de préparation et son application |
PCT/CN2023/092958 WO2023217125A1 (fr) | 2022-05-10 | 2023-05-09 | Cellule de type précurseur de la prostate et préparation cellulaire, procédé de préparation associé et utilisation associée |
Family Applications After (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2023/092967 WO2023217132A1 (fr) | 2022-05-10 | 2023-05-09 | Procédé de préparation et utilisation d'une cellule de type précurseur de vésicule biliaire |
PCT/CN2023/092956 WO2023217123A1 (fr) | 2022-05-10 | 2023-05-09 | Procédé de préparation et utilisation d'une cellule de type précurseur pulmonaire |
PCT/CN2023/092969 WO2023217134A1 (fr) | 2022-05-10 | 2023-05-09 | Préparation biologique contenant une cellule de type précurseur myocardique, son procédé de préparation et application |
PCT/CN2023/092964 WO2023217129A1 (fr) | 2022-05-10 | 2023-05-09 | Cellule de type précurseur de conduit biliaire intrahépatique, préparation cellulaire, procédé de préparation et application |
Country Status (2)
Country | Link |
---|---|
CN (9) | CN117018033A (fr) |
WO (9) | WO2023217126A1 (fr) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115948331A (zh) * | 2022-10-17 | 2023-04-11 | 中国人民解放军西部战区总医院 | 一种从小鼠胆囊中分离获取间充质干细胞的方法 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20170304370A1 (en) * | 2014-12-18 | 2017-10-26 | Pluristem Ltd. | Methods and compositions for treating and preventing muscle wasting disorders |
US20190338251A1 (en) * | 2016-04-27 | 2019-11-07 | Takeda Pharmaceutical Company Limited | Skeletal Muscle Precursor Cells and Production Method for Skeletal Muscle Cells |
CN113151150A (zh) * | 2020-04-09 | 2021-07-23 | 上海赛立维生物科技有限公司 | 胰腺细胞的扩增和分化方法以及应用 |
US20210244770A1 (en) * | 2018-05-08 | 2021-08-12 | Universität Zürich | Method for xeno-free generation of a population of hmpc |
CN113923990A (zh) * | 2019-04-03 | 2022-01-11 | 约翰·霍普金斯大学 | 产生骨骼肌干细胞和治疗疾病的方法、组合物和试剂盒 |
Family Cites Families (33)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010057013A1 (fr) * | 2008-11-14 | 2010-05-20 | Wake Forest University Health Sciences | Therapie cellulaire selective pour le traitement d'insuffisance renale |
CA2751332C (fr) * | 2009-02-03 | 2024-04-02 | Koninklijke Nederlandse Akademie Van Wetenschappen | Milieu de culture pour cellules souches epitheliales et organoides comprenant lesdites cellules souches |
CN102647989A (zh) * | 2009-10-19 | 2012-08-22 | 特里施泰姆贸易(塞浦路斯)有限公司 | 采用重编程序的成熟成体细胞的治疗 |
CA2817712C (fr) * | 2010-11-12 | 2020-03-24 | Georgetown University | Immortalisation de cellules epitheliales et leurs procedes d'utilisation |
WO2013049856A2 (fr) * | 2011-09-30 | 2013-04-04 | Hare Joshue M | Cellules souches rénales isolées du rein |
CN103031270A (zh) * | 2013-01-05 | 2013-04-10 | 绍兴文理学院 | 胆管上皮细胞的高效扩增和培养方法 |
CN104152398A (zh) * | 2013-05-13 | 2014-11-19 | 百奥迈科生物技术有限公司 | ***组织分离和建立原代上皮细胞和/或***的方法 |
ITFI20130303A1 (it) * | 2013-12-24 | 2015-06-25 | Azienda Ospedaliero Universitaria M Eyer | Metodica per l¿isolamento, purificazione ed amplificazione di progenitori renali cd133+cd24+ dalle urine di pazienti affetti da malattie renali. |
CN103860597A (zh) * | 2014-03-03 | 2014-06-18 | 奥思达干细胞有限公司 | 一种治疗缺血性心肌病的干细胞制剂及其制备方法 |
US9687512B2 (en) * | 2014-03-12 | 2017-06-27 | Banner Health | Isolated cardiac stem cells and methods of their use |
WO2015166638A1 (fr) * | 2014-05-01 | 2015-11-05 | iHeart Japan株式会社 | Cellules progénitrices cardiaques positives pour le cd82 |
SG10201902872YA (en) * | 2015-04-03 | 2019-04-29 | Propagenix Inc | Ex vivo proliferation of epithelial cells |
KR101782488B1 (ko) * | 2015-05-19 | 2017-09-28 | 주식회사 스템랩 | Oct4가 도입된 인간체세포로부터 직접적 리프로그래밍을 통한 희소돌기아교 전구세포를 유도하는 방법 |
CN104928235A (zh) * | 2015-07-10 | 2015-09-23 | 奥思达干细胞有限公司 | 基于干细胞的组合物及其在制备用于冠心病的制剂中的应用 |
DK3347450T3 (da) * | 2015-09-11 | 2021-05-31 | Propagenix Inc | Ex vivo-proliferation af epithelceller |
CN106754636B (zh) * | 2015-11-19 | 2019-08-30 | 中国人民解放军第二军医大学 | 体外诱导原代肝细胞胆管化并长期培养、扩增和分化的方法及其应用 |
JP2017108705A (ja) * | 2015-12-18 | 2017-06-22 | 国立大学法人京都大学 | 心筋細胞の製造方法 |
GB201603569D0 (en) * | 2016-03-01 | 2016-04-13 | Koninklijke Nederlandse Akademie Van Wetenschappen | Improved differentiation method |
CA3026318C (fr) * | 2016-06-01 | 2023-04-04 | Miltenyi Biotec Gmbh | Procede de production, d'identification et d'isolement de cardiomyocytes derives de cellules souches pluripotentes humaines et sous-populations de cardiomyocytes |
EP3483262B1 (fr) * | 2016-06-03 | 2021-11-17 | Institue of Transfusion Medicine, Academy of Military Medical Sciences, People's Liberation Army of China | Combinaison de composés à petites molécules pour reprogrammer des cellules épithéliales dérivées du tractus digestif en cellules souches/progénitrices endodermiques, procédé de reprogrammation et application |
CN107151645A (zh) * | 2017-05-16 | 2017-09-12 | 武汉大学深圳研究院 | 一种为肺癌提供离体个体化药物测试的方法及培养基 |
GB201709704D0 (en) * | 2017-06-19 | 2017-08-02 | Cambridge Entpr Ltd | Methods of expanding cholangiocytes |
CN108570443A (zh) * | 2017-12-05 | 2018-09-25 | 皓昇莱生物制药有限公司 | 一种用于培养尿液来源细胞的培养基 |
CN110218696A (zh) * | 2018-03-01 | 2019-09-10 | 中国科学院广州生物医药与健康研究院 | 一种用于化学诱导多能性干细胞生成的培养体系以及使用该培养体系的化学重编程方法 |
US20210238257A1 (en) * | 2018-04-27 | 2021-08-05 | The Regents Of The University Of California | De Novo Formation of the Biliary System by Hepatocyte Transdifferentiation |
KR102071302B1 (ko) * | 2018-06-28 | 2020-01-30 | 한국과학기술연구원 | 엑소좀 기반의 심근세포 교차분화 유도방법 |
CN109576215A (zh) * | 2018-12-27 | 2019-04-05 | 广州赛莱拉干细胞科技股份有限公司 | 一种诱导牙髓干细胞向心肌样细胞分化的方法 |
KR102150489B1 (ko) * | 2019-04-09 | 2020-09-01 | 고려대학교 산학협력단 | 소변세포로부터 신장전구세포로의 직접 역분화를 유도하는 방법 및 이의 방법으로 역분화된 신장전구세포를 포함하는 신장세포 손상 질환 예방 또는 치료용 약학 조성물 |
CN110184299B (zh) * | 2019-04-23 | 2023-11-24 | 中国科学院广州生物医药与健康研究院 | 诱导体细胞重编程的因子以及使用该因子诱导体细胞重编程的方法 |
CN112126618B (zh) * | 2019-06-24 | 2023-09-12 | 上海拜羡生物科技有限公司 | 一种人胆囊干细胞的获取和长期体外培养的方法 |
WO2021047495A1 (fr) * | 2019-09-12 | 2021-03-18 | 海门雨霖细胞科技有限责任公司 | Composition chimique à petites molécules et procédé pour induire chimiquement in vivo et in vitro la reprogrammation directe de fibroblastes en hépatocytes |
CN115322947B (zh) * | 2022-06-25 | 2024-02-09 | 同济大学 | 一种采用成体肾前体细胞经悬浮分化培养构建肾微器官的方法和应用 |
CN115992090A (zh) * | 2022-11-09 | 2023-04-21 | 同济大学 | 一种肾前体细胞来源的分泌组及其制备方法和应用 |
-
2023
- 2023-05-09 WO PCT/CN2023/092959 patent/WO2023217126A1/fr unknown
- 2023-05-09 CN CN202310522477.5A patent/CN117018033A/zh active Pending
- 2023-05-09 WO PCT/CN2023/092963 patent/WO2023217128A1/fr unknown
- 2023-05-09 CN CN202310522433.2A patent/CN117025507A/zh active Pending
- 2023-05-09 CN CN202310522470.3A patent/CN117025508A/zh active Pending
- 2023-05-09 WO PCT/CN2023/092954 patent/WO2023217121A1/fr unknown
- 2023-05-09 WO PCT/CN2023/092958 patent/WO2023217125A1/fr unknown
- 2023-05-09 CN CN202310519740.5A patent/CN117025503A/zh active Pending
- 2023-05-09 WO PCT/CN2023/092965 patent/WO2023217130A1/fr unknown
- 2023-05-09 CN CN202310522452.5A patent/CN117018032A/zh active Pending
- 2023-05-09 WO PCT/CN2023/092967 patent/WO2023217132A1/fr unknown
- 2023-05-09 WO PCT/CN2023/092956 patent/WO2023217123A1/fr unknown
- 2023-05-09 CN CN202310519743.9A patent/CN117025504A/zh active Pending
- 2023-05-09 CN CN202310519744.3A patent/CN117025505A/zh active Pending
- 2023-05-09 CN CN202310519697.2A patent/CN117025502A/zh active Pending
- 2023-05-09 WO PCT/CN2023/092969 patent/WO2023217134A1/fr unknown
- 2023-05-09 CN CN202310522421.XA patent/CN117025506A/zh active Pending
- 2023-05-09 WO PCT/CN2023/092964 patent/WO2023217129A1/fr unknown
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20170304370A1 (en) * | 2014-12-18 | 2017-10-26 | Pluristem Ltd. | Methods and compositions for treating and preventing muscle wasting disorders |
US20190338251A1 (en) * | 2016-04-27 | 2019-11-07 | Takeda Pharmaceutical Company Limited | Skeletal Muscle Precursor Cells and Production Method for Skeletal Muscle Cells |
US20210244770A1 (en) * | 2018-05-08 | 2021-08-12 | Universität Zürich | Method for xeno-free generation of a population of hmpc |
CN113923990A (zh) * | 2019-04-03 | 2022-01-11 | 约翰·霍普金斯大学 | 产生骨骼肌干细胞和治疗疾病的方法、组合物和试剂盒 |
CN113151150A (zh) * | 2020-04-09 | 2021-07-23 | 上海赛立维生物科技有限公司 | 胰腺细胞的扩增和分化方法以及应用 |
Non-Patent Citations (2)
Title |
---|
FU XIN, XIAO JUN, WEI YUNING, LI SHENG, LIU YAN, YIN JIE, SUN KUN, SUN HAO, WANG HUATING, ZHANG ZONGKANG, ZHANG BAO-TING, SHENG CH: "Combination of inflammation-related cytokines promotes long-term muscle stem cell expansion", CELL RESEARCH, SPRINGER SINGAPORE, SINGAPORE, vol. 25, no. 6, 1 June 2015 (2015-06-01), Singapore , pages 655 - 673, XP093109144, ISSN: 1001-0602, DOI: 10.1038/cr.2015.58 * |
YANG WENJUN; HU PING: "Current Progress on Cell Replacement Therapy", CHINESE JOURNAL OF CELL BIOLOGY, ZHONGGUO KEXUEYUAN SHANGHAI SHENGMING KEXUE YANJIUYUAN SHENGWU HUAXUE YU XIBAO SHENGWUXUE YANJIUSUO, CN, vol. 41, no. 4, 9 May 2019 (2019-05-09), CN , pages 549 - 560, XP009550440, ISSN: 1674-7666 * |
Also Published As
Publication number | Publication date |
---|---|
CN117018033A (zh) | 2023-11-10 |
CN117025507A (zh) | 2023-11-10 |
WO2023217126A1 (fr) | 2023-11-16 |
CN117025508A (zh) | 2023-11-10 |
WO2023217123A1 (fr) | 2023-11-16 |
CN117025503A (zh) | 2023-11-10 |
WO2023217121A1 (fr) | 2023-11-16 |
WO2023217134A1 (fr) | 2023-11-16 |
CN117025502A (zh) | 2023-11-10 |
CN117018032A (zh) | 2023-11-10 |
CN117025506A (zh) | 2023-11-10 |
CN117025505A (zh) | 2023-11-10 |
CN117025504A (zh) | 2023-11-10 |
WO2023217129A1 (fr) | 2023-11-16 |
WO2023217128A1 (fr) | 2023-11-16 |
WO2023217125A1 (fr) | 2023-11-16 |
WO2023217132A1 (fr) | 2023-11-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
ES2373551T3 (es) | Métodos para usar células derivadas de tejido adiposo en el tratamiento de afecciones cardiovasculares. | |
US20210145893A1 (en) | Compositions to amplify cardiac stem cells in vitro and in viv | |
CN112760293B (zh) | 一种无异源血清3d培养msc干细胞制备高活性外泌体的方法 | |
CN108348555B (zh) | 细胞扩增方法和治疗组合物 | |
WO2010069204A1 (fr) | Cellules souches pluripotentes, procédé pour la préparation de celles-ci et utilisations de celles-ci | |
CN106916783B (zh) | 肌肉干细胞体外培养方法及其应用 | |
WO2015137419A1 (fr) | Activateur de cellules souches mésenchymateuses, cellules mésenchymateuses activées, et leur procédé de production | |
WO2023217130A1 (fr) | Formulation biologique contenant des cellules de type précurseur de muscle squelettique, son procédé de préparation et son application | |
Teng et al. | Treatment of acute hepatic failure in mice by transplantation of mixed microencapsulation of rat hepatocytes and transgenic human fetal liver stromal cells | |
WO2021147923A1 (fr) | Vésicule et utilisation de celle-ci | |
EP4394031A1 (fr) | Cellule souche mésenchymateuse et son exosome obtenus par traitement d'au moins deux cytokines parmi il4, il21 et il27, et utilisation associée | |
Chen et al. | Directional homing of glycosylation-modified bone marrow mesenchymal stem cells for bone defect repair | |
WO2006134951A1 (fr) | Procédé de prolifération de culture à long terme pour cellule souche dérivée de graisse | |
CN106606512B (zh) | 一种用于治疗心肌梗死的混合细胞制剂及制备方法与应用 | |
CN114177203A (zh) | 脐带血浓缩细胞治疗卵巢早衰的细胞治疗剂 | |
CN111973632B (zh) | 一种用于治疗糖尿病的干细胞制剂及其制备方法 | |
Hong | Fat grafts enriched with adipose-derived stem cells | |
WO2014201986A1 (fr) | Procédé de production de cellule souche mésenchymateuse monoclonale et utilisation correspondante | |
WO2024045404A1 (fr) | Surnageant de moelle osseuse et son utilisation dans la culture cellulaire | |
CN113073078B (zh) | 脐带来源间充质干细胞制备的通用型血小板制剂及方法 | |
EP2205251B1 (fr) | Procédé pour amplifier des cellules cardiaques souches in vitro et in vivo | |
WO2020190672A1 (fr) | Exosomes dérivés de cardiomyocytes induisant la régénération de tissu cardiaque endommagé | |
CN111514164A (zh) | 一种用于肺病治疗的脂肪间充质干细胞及其制备方法 | |
CN118240883B (zh) | 一种炎症调节能力增强型脐带间充质干细胞及其应用、制备方法 | |
WO2017197648A1 (fr) | Composition pharmaceutique destinée au traitement de la cardiopathie |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 23802899 Country of ref document: EP Kind code of ref document: A1 |