JP7387115B2 - cP1Pまたはその薬学的に許容可能な塩を有効成分として含む幹細胞増殖促進用組成物 - Google Patents
cP1Pまたはその薬学的に許容可能な塩を有効成分として含む幹細胞増殖促進用組成物 Download PDFInfo
- Publication number
- JP7387115B2 JP7387115B2 JP2022500607A JP2022500607A JP7387115B2 JP 7387115 B2 JP7387115 B2 JP 7387115B2 JP 2022500607 A JP2022500607 A JP 2022500607A JP 2022500607 A JP2022500607 A JP 2022500607A JP 7387115 B2 JP7387115 B2 JP 7387115B2
- Authority
- JP
- Japan
- Prior art keywords
- stem cell
- cp1p
- composition
- stem cells
- cells
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 210000000130 stem cell Anatomy 0.000 title claims description 165
- 239000000203 mixture Substances 0.000 title claims description 66
- 230000004663 cell proliferation Effects 0.000 title claims description 64
- 230000001737 promoting effect Effects 0.000 title claims description 49
- 150000003839 salts Chemical class 0.000 title claims description 30
- 239000004480 active ingredient Substances 0.000 title claims description 16
- 210000001671 embryonic stem cell Anatomy 0.000 claims description 26
- 238000012258 culturing Methods 0.000 claims description 24
- 210000004263 induced pluripotent stem cell Anatomy 0.000 claims description 24
- AYGOSKULTISFCW-KSZLIROESA-N phytosphingosine 1-phosphate Chemical compound CCCCCCCCCCCCCC[C@@H](O)[C@@H](O)[C@@H](N)COP(O)(O)=O AYGOSKULTISFCW-KSZLIROESA-N 0.000 claims description 24
- 230000006907 apoptotic process Effects 0.000 claims description 21
- 238000000034 method Methods 0.000 claims description 20
- 239000006143 cell culture medium Substances 0.000 claims description 18
- 238000000338 in vitro Methods 0.000 claims description 17
- 210000002966 serum Anatomy 0.000 claims description 16
- 238000004113 cell culture Methods 0.000 claims description 10
- 230000034994 death Effects 0.000 claims description 5
- 210000004027 cell Anatomy 0.000 description 68
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 42
- 230000001965 increasing effect Effects 0.000 description 30
- 230000014509 gene expression Effects 0.000 description 20
- 230000035755 proliferation Effects 0.000 description 20
- 238000011282 treatment Methods 0.000 description 16
- 210000001778 pluripotent stem cell Anatomy 0.000 description 15
- 108090000623 proteins and genes Proteins 0.000 description 15
- DUYSYHSSBDVJSM-KRWOKUGFSA-N sphingosine 1-phosphate Chemical compound CCCCCCCCCCCCC\C=C\[C@@H](O)[C@@H](N)COP(O)(O)=O DUYSYHSSBDVJSM-KRWOKUGFSA-N 0.000 description 15
- 239000002609 medium Substances 0.000 description 13
- 239000001963 growth medium Substances 0.000 description 12
- 150000001875 compounds Chemical class 0.000 description 9
- 210000001519 tissue Anatomy 0.000 description 9
- 241001465754 Metazoa Species 0.000 description 8
- 230000030833 cell death Effects 0.000 description 8
- 230000000694 effects Effects 0.000 description 8
- 230000022131 cell cycle Effects 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- 101710126211 POU domain, class 5, transcription factor 1 Proteins 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- 238000012790 confirmation Methods 0.000 description 6
- 230000004069 differentiation Effects 0.000 description 6
- 239000012453 solvate Substances 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 230000003247 decreasing effect Effects 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 238000011866 long-term treatment Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 102000004121 Annexin A5 Human genes 0.000 description 4
- 108090000672 Annexin A5 Proteins 0.000 description 4
- 102000000905 Cadherin Human genes 0.000 description 4
- 108050007957 Cadherin Proteins 0.000 description 4
- 102100035423 POU domain, class 5, transcription factor 1 Human genes 0.000 description 4
- 239000000872 buffer Substances 0.000 description 4
- -1 cP1P Chemical compound 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000003013 cytotoxicity Effects 0.000 description 4
- 231100000135 cytotoxicity Toxicity 0.000 description 4
- 230000007774 longterm Effects 0.000 description 4
- 239000012679 serum free medium Substances 0.000 description 4
- OZFAFGSSMRRTDW-UHFFFAOYSA-N (2,4-dichlorophenyl) benzenesulfonate Chemical compound ClC1=CC(Cl)=CC=C1OS(=O)(=O)C1=CC=CC=C1 OZFAFGSSMRRTDW-UHFFFAOYSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 239000012591 Dulbecco’s Phosphate Buffered Saline Substances 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 241000282412 Homo Species 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- 239000012148 binding buffer Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000010494 dissociation reaction Methods 0.000 description 3
- 230000005593 dissociations Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 230000036542 oxidative stress Effects 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- YXHLJMWYDTXDHS-IRFLANFNSA-N 7-aminoactinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=C(N)C=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 YXHLJMWYDTXDHS-IRFLANFNSA-N 0.000 description 2
- 108700012813 7-aminoactinomycin D Proteins 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 102000002427 Cyclin B Human genes 0.000 description 2
- 108010068150 Cyclin B Proteins 0.000 description 2
- 102100032857 Cyclin-dependent kinase 1 Human genes 0.000 description 2
- 101710106279 Cyclin-dependent kinase 1 Proteins 0.000 description 2
- RGHNJXZEOKUKBD-SQOUGZDYSA-N D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- 239000006147 Glasgow's Minimal Essential Medium Substances 0.000 description 2
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 description 2
- 101000687905 Homo sapiens Transcription factor SOX-2 Proteins 0.000 description 2
- 101000976622 Homo sapiens Zinc finger protein 42 homolog Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 108700021430 Kruppel-Like Factor 4 Proteins 0.000 description 2
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 2
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 101100247004 Rattus norvegicus Qsox1 gene Proteins 0.000 description 2
- 108700005075 Regulator Genes Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 102100024270 Transcription factor SOX-2 Human genes 0.000 description 2
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 210000004504 adult stem cell Anatomy 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000001185 bone marrow Anatomy 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 230000005754 cellular signaling Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000012531 culture fluid Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 235000013601 eggs Nutrition 0.000 description 2
- 210000002242 embryoid body Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000010195 expression analysis Methods 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- 238000010185 immunofluorescence analysis Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 150000007522 mineralic acids Chemical class 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000013630 prepared media Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 239000004017 serum-free culture medium Substances 0.000 description 2
- 210000001082 somatic cell Anatomy 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 210000003014 totipotent stem cell Anatomy 0.000 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- RTBFRGCFXZNCOE-UHFFFAOYSA-N 1-methylsulfonylpiperidin-4-one Chemical compound CS(=O)(=O)N1CCC(=O)CC1 RTBFRGCFXZNCOE-UHFFFAOYSA-N 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 201000004384 Alopecia Diseases 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 101150069146 C5 gene Proteins 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- 101150027068 DEGS1 gene Proteins 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 230000004668 G2/M phase Effects 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- 238000003559 RNA-seq method Methods 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 206010038997 Retroviral infections Diseases 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 102100035140 Vitronectin Human genes 0.000 description 1
- 108010031318 Vitronectin Proteins 0.000 description 1
- 235000011054 acetic acid Nutrition 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- JFCQEDHGNNZCLN-UHFFFAOYSA-N anhydrous glutaric acid Natural products OC(=O)CCCC(O)=O JFCQEDHGNNZCLN-UHFFFAOYSA-N 0.000 description 1
- 230000001640 apoptogenic effect Effects 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 239000007640 basal medium Substances 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 210000002459 blastocyst Anatomy 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
- SXDBWCPKPHAZSM-UHFFFAOYSA-N bromic acid Chemical compound OBr(=O)=O SXDBWCPKPHAZSM-UHFFFAOYSA-N 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 235000011148 calcium chloride Nutrition 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 230000011712 cell development Effects 0.000 description 1
- 238000002659 cell therapy Methods 0.000 description 1
- 230000033077 cellular process Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 210000002257 embryonic structure Anatomy 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 235000011087 fumaric acid Nutrition 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000000174 gluconic acid Substances 0.000 description 1
- 235000012208 gluconic acid Nutrition 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000003966 growth inhibitor Substances 0.000 description 1
- 230000003779 hair growth Effects 0.000 description 1
- 208000024963 hair loss Diseases 0.000 description 1
- 230000003676 hair loss Effects 0.000 description 1
- 210000003566 hemangioblast Anatomy 0.000 description 1
- 150000004677 hydrates Chemical class 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 239000002829 mitogen activated protein kinase inhibitor Substances 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 230000001817 pituitary effect Effects 0.000 description 1
- 210000002826 placenta Anatomy 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000002250 progressing effect Effects 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 230000008672 reprogramming Effects 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- YXFVVABEGXRONW-UHFFFAOYSA-N toluene Substances CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
- 210000003556 vascular endothelial cell Anatomy 0.000 description 1
- 210000004509 vascular smooth muscle cell Anatomy 0.000 description 1
- 238000001262 western blot Methods 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
- C12N5/0603—Embryonic cells ; Embryoid bodies
- C12N5/0606—Pluripotent embryonic cells, e.g. embryonic stem cells [ES]
-
- 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
- C12N5/0696—Artificially induced pluripotent stem cells, e.g. iPS
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/36—Lipids
-
- 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.)
-
- 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
- C12N2501/999—Small molecules not provided for elsewhere
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Reproductive Health (AREA)
- Gynecology & Obstetrics (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- Transplantation (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Description
cP1P(o-cyclic phytosphingosine-1-phosphate)濃度によるhPSCs増殖能の変化を確認するため、hPSCsを培養し、cP1Pを濃度別に処理した後、hPSCs増殖の変化を確認した。
hPSCs増殖能を示すcP1Pの最適濃度を決定するために、1-500nM濃度において細胞増殖能を確認した。
cP1P処理によるhPSCsの分化能変化の有無を確認するために、実施例2と同様の方法で10nM cP1PをhPSC細胞に処理したり、或いは対照群(vehicle,DMSO)を処理した後、アルカリンホスファターゼ染色キット(Sigma-Aldrich US,SCR004)を用いて染色した後、光学顕微鏡で観察した。
cP1P処理によるコロニー数及び大きさの変化を以下のような方法で確認した。具体的には、実施例1と同様の方法でhPSCsに1nM、10nM、100nM、1,000nM、または10,000nMのcP1Pを処理し、6日間培養し、7日目に光学顕微鏡下でコロニー数および大きさを確認した。
その結果、図5に示すように、10nM以上のcP1PをhPSCsに処理した場合、対照群に比べて大きさが150mm以上のコロニー数が有意に増加した(P<0.01)。
cP1P処理によるhPSCs増殖能の変化が継代培養時、持続するか否かを確認するために、実施例1と同様の方法で10nM cP1PをhPSCsに持続的に処理しながら3回の継代培養を行った後、hPSCs細胞増殖能の変化を確認した。
cP1P処理によるhPSCs細胞周期の変化を確認するために、フローサイトメトリー解析(flow cytometer)、アポトーシス解析、及び細胞周期調節遺伝子の発現解析を行った。
cP1PをhPSCsに処理する場合、アポトーシスおよび細胞毒性が誘導されるかどうかを確認するために、実施例6と同様の方法で10nMまたは100nM cP1PをhPSCsに5日間処理した後、Annexin V apoptosis detection kit(BD Pharmingen US,556547)を用いて解析した。
cP1P処理によってhPSCsの全能性特性が増加するか否かを以下の方法で確認した。
ヒトPSCsを長期培養する場合、増殖速度が減少したり、幹細胞の性質が微細に変わるなどの変化をもたらしたりするため、hPSCsにcP1Pを長期的に処理する場合、hPSCsの特性に変化が発生するかどうかを確認した。
Claims (13)
- cP1P(o-cyclic phytosphingosine-1-phosphate)またはその薬学的に許容可能な塩を有効成分として含む幹細胞増殖促進用組成物であって、
前記幹細胞は、胚性幹細胞(Embryonic stem cell)または人工多能性幹細胞(induced Pluripotent stem cell)である、幹細胞増殖促進用組成物。 - 前記cP1Pは、0.1nM~10,000nM濃度であることを特徴とする、請求項1に記載の幹細胞増殖促進用組成物。
- 前記組成物は、幹細胞の増殖を促進し、アポトーシスを阻害することを特徴とする、請求項1に記載の幹細胞増殖促進用組成物。
- 前記組成物は、幹細胞コロニーの数を増加させ、コロニーの大きさを増加させることを特徴とする、請求項1に記載の幹細胞増殖促進用組成物。
- 前記組成物は、幹細胞の全能性(stemness, naive state)を強化することを特徴とする、請求項1に記載の幹細胞増殖促進用組成物。
- 前記幹細胞培養用組成物は、無血清または血清成分を0.1~3重量%含有することを特徴とする、請求項1に記載の幹細胞増殖促進用組成物。
- 幹細胞培養物に添加する組成物であって、前記組成物はcP1Pまたはその薬学的に許容可能な塩を有効成分として含み、
前記幹細胞は、胚性幹細胞(Embryonic stem cell)または人工多能性幹細胞(induced Pluripotent stem cell)である、幹細胞培養液添加用組成物。 - 前記cP1Pは、0.1nM~10,000nM濃度であることを特徴とする、請求項7に記載の幹細胞培養液添加用組成物。
- 請求項1に記載の組成物を幹細胞に処理して培養する段階を含む、幹細胞培養方法であって、
前記幹細胞は胚性幹細胞(Embryonic stem cell)または人工多能性幹細胞(induced Pluripotent stem cell)である、幹細胞培養方法。 - 請求項1に記載の組成物を含む、体外培養環境における幹細胞死滅抑制用キットであって、
前記幹細胞は胚性幹細胞(Embryonic stem cell)または人工多能性幹細胞(induced Pluripotent stem cell)である、キット。 - 請求項1に記載の組成物を含む、体外培養環境における幹細胞増殖促進用キットであって、
前記幹細胞は胚性幹細胞(Embryonic stem cell)または人工多能性幹細胞(induced Pluripotent stem cell)である、キット。 - cP1P(o-cyclic phytosphingosine-1-phosphate)またはその薬学的に許容可能な塩を幹細胞培養物に添加して幹細胞を培養する段階を含む、幹細胞増殖促進方法であって、
前記幹細胞は胚性幹細胞(Embryonic stem cell)または人工多能性幹細胞(induced Pluripotent stem cell)である、幹細胞増殖促進方法。 - 前記cP1Pは、0.1nM~10,000nM濃度であることを特徴とする、請求項12に記載の幹細胞増殖促進方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR10-2019-0079547 | 2019-07-02 | ||
KR1020190079547A KR102276781B1 (ko) | 2019-07-02 | 2019-07-02 | cP1P 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 줄기세포 증식 촉진용 조성물 |
PCT/KR2020/001037 WO2021002554A1 (ko) | 2019-07-02 | 2020-01-21 | Cp1p 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 줄기세포 증식 촉진용 조성물 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2022540579A JP2022540579A (ja) | 2022-09-16 |
JP7387115B2 true JP7387115B2 (ja) | 2023-11-28 |
Family
ID=74101089
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2022500607A Active JP7387115B2 (ja) | 2019-07-02 | 2020-01-21 | cP1Pまたはその薬学的に許容可能な塩を有効成分として含む幹細胞増殖促進用組成物 |
Country Status (6)
Country | Link |
---|---|
US (1) | US20220380724A1 (ja) |
EP (1) | EP3995570A4 (ja) |
JP (1) | JP7387115B2 (ja) |
KR (1) | KR102276781B1 (ja) |
CN (1) | CN114127264A (ja) |
WO (1) | WO2021002554A1 (ja) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20220166662A (ko) * | 2021-06-10 | 2022-12-19 | 한양대학교 산학협력단 | 줄기세포를 혈관내피세포로 분화 유도하기 위한 배지 조성물 |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US556422A (en) | 1896-03-17 | Machines | ||
US556547A (en) | 1896-03-17 | Arthur marichal | ||
US1559925A (en) | 1923-09-20 | 1925-11-03 | Goodrich Co B F | Composition of matter and method of producing the same |
WO2012134134A2 (ko) * | 2011-03-25 | 2012-10-04 | (주)다미화학 | 피토스핑고신-1-포스페이트를 유효성분으로 포함하는 화장료 조성물 |
KR101340556B1 (ko) * | 2012-02-16 | 2013-12-11 | 주식회사 피토스 | 신규한 파이토스핑고신-1-포스페이트 유도체, 그 제조방법, 및 그것을 포함하는 탈모의 예방, 치료, 또는 육모용 조성물 |
EP2982678B1 (en) * | 2013-04-04 | 2017-09-20 | Phytos Co., Ltd. | Novel phytospingosine-1-phosphate derivative, preparation method therefor, and composition for preventing and treating hair loss or for growing hair comprising same |
KR101514970B1 (ko) * | 2013-08-28 | 2015-04-24 | 주식회사 피토스 | 피토스핑고신-1-포스페이트 또는 그 유도체를 포함하는 아토피 또는 피부상처 치료 또는 예방용 조성물 |
KR101900818B1 (ko) * | 2016-05-17 | 2018-09-20 | 주식회사 피토스 | 피토스핑고신-1-포스페이트 또는 그 유도체를 포함하는 줄기세포 성장 촉진용 조성물 및 이를 포함하는 줄기세포 배양배지용 조성물 |
KR101965729B1 (ko) * | 2018-08-23 | 2019-04-03 | 주식회사 엑세쏘바이오파마 | 피토스핑고신-1-포스페이트 유도체를 포함하는 줄기세포 성장 촉진용 조성물 및 이를 포함하는 줄기세포 배양배지용 조성물 |
-
2019
- 2019-07-02 KR KR1020190079547A patent/KR102276781B1/ko active IP Right Grant
-
2020
- 2020-01-21 JP JP2022500607A patent/JP7387115B2/ja active Active
- 2020-01-21 CN CN202080049187.8A patent/CN114127264A/zh active Pending
- 2020-01-21 US US17/624,434 patent/US20220380724A1/en active Pending
- 2020-01-21 EP EP20835482.9A patent/EP3995570A4/en active Pending
- 2020-01-21 WO PCT/KR2020/001037 patent/WO2021002554A1/ko unknown
Also Published As
Publication number | Publication date |
---|---|
JP2022540579A (ja) | 2022-09-16 |
EP3995570A4 (en) | 2023-07-26 |
US20220380724A1 (en) | 2022-12-01 |
WO2021002554A1 (ko) | 2021-01-07 |
EP3995570A1 (en) | 2022-05-11 |
KR20210003574A (ko) | 2021-01-12 |
KR102276781B1 (ko) | 2021-07-14 |
CN114127264A (zh) | 2022-03-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5227318B2 (ja) | 細胞増殖培地 | |
JP5479661B2 (ja) | 幹細胞の分化を誘導する方法 | |
JP2020115866A (ja) | 多能性幹細胞から機能的心筋へと直接分化させる方法 | |
EP1983042B1 (en) | A method for purifying cardiomyocytes or programmed cardiomyocytes derived from stem cells or fetuses | |
US20080254003A1 (en) | Differentiation of Human Embryonic Stem Cells and Cardiomyocytes and Cardiomyocyte Progenitors Derived Therefrom | |
US20040120932A1 (en) | In vitro-derived adult pluripotent stem cells and uses therefor | |
AU2002317039A1 (en) | Methods of inducing differentiation of stem cells | |
JP2017532047A (ja) | 多能性幹細胞由来ケラチノサイトの生成およびケラチノサイト培養の維持 | |
US20130302887A1 (en) | Compositions and methods for growing human embryonic cells | |
JP2023052609A (ja) | 細胞培養物の製造方法 | |
KR20220016846A (ko) | 만능세포 응집체 및 그의 용도 | |
JP7387115B2 (ja) | cP1Pまたはその薬学的に許容可能な塩を有効成分として含む幹細胞増殖促進用組成物 | |
Yamasaki et al. | Long-term serial cultivation of mouse induced pluripotent stem cells in serum-free and feeder-free defined medium. | |
WO2020203532A1 (ja) | 多能性幹細胞の製造方法 | |
JP2008500821A (ja) | 心筋細胞への分化の改善 | |
EP4317418A1 (en) | Method for producing ovarian somatic cell-like cells, and method for inducing differentiation of primate pluripotent stem cells into ovarian somatic cell-like cells | |
KR102180598B1 (ko) | 줄기세포를 심근세포로 분화 유도하기 위한 배지 조성물 | |
JP2022504764A (ja) | 細胞培養のための組成物および方法 | |
KR101177869B1 (ko) | 옥트(Oct)-4 발현능을 가지는 피부 유래 다분화능 성체줄기세포 및 그의 제조방법 | |
KR20190070254A (ko) | 페닐부틸산나트륨을 포함하는 고효율 세포전환용 배지 첨가제 | |
KR20060114381A (ko) | 배아줄기세포용 피더세포 작성 배지 및 피더세포 | |
AU2005318931A1 (en) | Differentiation of human embryonic stem cells and cardiomyocytes and cardiomyocyte progenitors derived therefrom | |
Rajala | Development of human stem cell culture conditions for clinical cell therapy |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20220414 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20230404 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20230704 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20230904 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20230904 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20231010 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20231106 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 7387115 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |