JP6307515B2 - 肝細胞機能の増強のための細胞培養培地 - Google Patents
肝細胞機能の増強のための細胞培養培地 Download PDFInfo
- Publication number
- JP6307515B2 JP6307515B2 JP2015544169A JP2015544169A JP6307515B2 JP 6307515 B2 JP6307515 B2 JP 6307515B2 JP 2015544169 A JP2015544169 A JP 2015544169A JP 2015544169 A JP2015544169 A JP 2015544169A JP 6307515 B2 JP6307515 B2 JP 6307515B2
- Authority
- JP
- Japan
- Prior art keywords
- cells
- liver
- lfm
- composition
- cultured
- 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
- 210000003494 hepatocyte Anatomy 0.000 title claims description 92
- 239000006143 cell culture medium Substances 0.000 title description 21
- 150000001875 compounds Chemical class 0.000 claims description 92
- 239000000203 mixture Substances 0.000 claims description 83
- 210000004027 cell Anatomy 0.000 claims description 79
- 210000005229 liver cell Anatomy 0.000 claims description 73
- UVSVTDVJQAJIFG-VURMDHGXSA-N LFM-A13 Chemical compound C\C(O)=C(/C#N)C(=O)NC1=CC(Br)=CC=C1Br UVSVTDVJQAJIFG-VURMDHGXSA-N 0.000 claims description 56
- 102000004190 Enzymes Human genes 0.000 claims description 36
- 108090000790 Enzymes Proteins 0.000 claims description 36
- 238000000034 method Methods 0.000 claims description 36
- 229940079593 drug Drugs 0.000 claims description 23
- 239000003814 drug Substances 0.000 claims description 23
- 238000012258 culturing Methods 0.000 claims description 14
- 230000002708 enhancing effect Effects 0.000 claims description 12
- 239000002207 metabolite Substances 0.000 claims description 9
- 210000004748 cultured cell Anatomy 0.000 claims description 8
- 230000007102 metabolic function Effects 0.000 claims description 8
- 210000002966 serum Anatomy 0.000 claims description 5
- 230000000694 effects Effects 0.000 description 68
- VHOGYURTWQBHIL-UHFFFAOYSA-N leflunomide Chemical compound O1N=CC(C(=O)NC=2C=CC(=CC=2)C(F)(F)F)=C1C VHOGYURTWQBHIL-UHFFFAOYSA-N 0.000 description 39
- 108010081668 Cytochrome P-450 CYP3A Proteins 0.000 description 36
- 102100039205 Cytochrome P450 3A4 Human genes 0.000 description 30
- 210000000130 stem cell Anatomy 0.000 description 29
- 230000006870 function Effects 0.000 description 27
- 239000002609 medium Substances 0.000 description 27
- 210000004185 liver Anatomy 0.000 description 26
- 229960000681 leflunomide Drugs 0.000 description 23
- 230000036267 drug metabolism Effects 0.000 description 21
- 230000014509 gene expression Effects 0.000 description 21
- 230000003908 liver function Effects 0.000 description 19
- 108090000623 proteins and genes Proteins 0.000 description 19
- 102000002004 Cytochrome P-450 Enzyme System Human genes 0.000 description 18
- 108010015742 Cytochrome P-450 Enzyme System Proteins 0.000 description 17
- 238000004113 cell culture Methods 0.000 description 15
- 108020004999 messenger RNA Proteins 0.000 description 15
- -1 LFM Chemical compound 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 14
- 125000006273 (C1-C3) alkyl group Chemical group 0.000 description 12
- 108010074922 Cytochrome P-450 CYP1A2 Proteins 0.000 description 12
- 102100026533 Cytochrome P450 1A2 Human genes 0.000 description 12
- 102000040945 Transcription factor Human genes 0.000 description 12
- 108091023040 Transcription factor Proteins 0.000 description 12
- 108010088751 Albumins Proteins 0.000 description 11
- 102000009027 Albumins Human genes 0.000 description 11
- 230000004069 differentiation Effects 0.000 description 11
- 108020005497 Nuclear hormone receptor Proteins 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 9
- 230000001965 increasing effect Effects 0.000 description 9
- 102000006255 nuclear receptors Human genes 0.000 description 9
- 108020004017 nuclear receptors Proteins 0.000 description 9
- 108090000448 Aryl Hydrocarbon Receptors Proteins 0.000 description 8
- 102000003984 Aryl Hydrocarbon Receptors Human genes 0.000 description 8
- 102000012422 Collagen Type I Human genes 0.000 description 8
- 108010022452 Collagen Type I Proteins 0.000 description 8
- 238000011534 incubation Methods 0.000 description 8
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 8
- 229960001225 rifampicin Drugs 0.000 description 8
- 230000001225 therapeutic effect Effects 0.000 description 8
- 229940124291 BTK inhibitor Drugs 0.000 description 7
- 210000001671 embryonic stem cell Anatomy 0.000 description 7
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 7
- LMBFAGIMSUYTBN-MPZNNTNKSA-N teixobactin Chemical compound C([C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H](CCC(N)=O)C(=O)N[C@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H]1C(N[C@@H](C)C(=O)N[C@@H](C[C@@H]2NC(=N)NC2)C(=O)N[C@H](C(=O)O[C@H]1C)[C@@H](C)CC)=O)NC)C1=CC=CC=C1 LMBFAGIMSUYTBN-MPZNNTNKSA-N 0.000 description 7
- 102100028161 ATP-binding cassette sub-family C member 2 Human genes 0.000 description 6
- 108010092364 Glucuronosyltransferase Proteins 0.000 description 6
- 102000016354 Glucuronosyltransferase Human genes 0.000 description 6
- 102100022054 Hepatocyte nuclear factor 4-alpha Human genes 0.000 description 6
- 101001045740 Homo sapiens Hepatocyte nuclear factor 4-alpha Proteins 0.000 description 6
- 108010066419 Multidrug Resistance-Associated Protein 2 Proteins 0.000 description 6
- 102000034527 Retinoid X Receptors Human genes 0.000 description 6
- 108010038912 Retinoid X Receptors Proteins 0.000 description 6
- 108090001033 Sulfotransferases Proteins 0.000 description 6
- 102000004896 Sulfotransferases Human genes 0.000 description 6
- 238000001952 enzyme assay Methods 0.000 description 6
- 125000001475 halogen functional group Chemical group 0.000 description 6
- 239000001257 hydrogen Substances 0.000 description 6
- 229910052739 hydrogen Inorganic materials 0.000 description 6
- 230000003993 interaction Effects 0.000 description 6
- 210000001778 pluripotent stem cell Anatomy 0.000 description 6
- 102000005962 receptors Human genes 0.000 description 6
- 108020003175 receptors Proteins 0.000 description 6
- 208000008964 Chemical and Drug Induced Liver Injury Diseases 0.000 description 5
- 102000008169 Co-Repressor Proteins Human genes 0.000 description 5
- 108010060434 Co-Repressor Proteins Proteins 0.000 description 5
- 206010072268 Drug-induced liver injury Diseases 0.000 description 5
- 241000700605 Viruses Species 0.000 description 5
- 125000003118 aryl group Chemical group 0.000 description 5
- 238000003556 assay Methods 0.000 description 5
- 238000003149 assay kit Methods 0.000 description 5
- 239000003112 inhibitor Substances 0.000 description 5
- 239000003550 marker Substances 0.000 description 5
- 102100023635 Alpha-fetoprotein Human genes 0.000 description 4
- 108010078791 Carrier Proteins Proteins 0.000 description 4
- 102000004328 Cytochrome P-450 CYP3A Human genes 0.000 description 4
- 102100038637 Cytochrome P450 7A1 Human genes 0.000 description 4
- 101000582926 Dictyostelium discoideum Probable serine/threonine-protein kinase PLK Proteins 0.000 description 4
- 108090000079 Glucocorticoid Receptors Proteins 0.000 description 4
- 102100033417 Glucocorticoid receptor Human genes 0.000 description 4
- 108010070675 Glutathione transferase Proteins 0.000 description 4
- 102100029100 Hematopoietic prostaglandin D synthase Human genes 0.000 description 4
- 102000006752 Hepatocyte Nuclear Factor 4 Human genes 0.000 description 4
- 125000001589 carboacyl group Chemical group 0.000 description 4
- 230000003915 cell function Effects 0.000 description 4
- 150000003384 small molecules Chemical class 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 102000009310 vitamin D receptors Human genes 0.000 description 4
- 108050000156 vitamin D receptors Proteins 0.000 description 4
- 125000004191 (C1-C6) alkoxy group Chemical group 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 102000005416 ATP-Binding Cassette Transporters Human genes 0.000 description 3
- 108010006533 ATP-Binding Cassette Transporters Proteins 0.000 description 3
- 102000008130 Cyclic AMP-Dependent Protein Kinases Human genes 0.000 description 3
- 108010049894 Cyclic AMP-Dependent Protein Kinases Proteins 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 102100022057 Hepatocyte nuclear factor 1-alpha Human genes 0.000 description 3
- 102100029087 Hepatocyte nuclear factor 6 Human genes 0.000 description 3
- 101001045751 Homo sapiens Hepatocyte nuclear factor 1-alpha Proteins 0.000 description 3
- 101000988619 Homo sapiens Hepatocyte nuclear factor 6 Proteins 0.000 description 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- ORNBQBCIOKFOEO-YQUGOWONSA-N Pregnenolone Natural products O=C(C)[C@@H]1[C@@]2(C)[C@H]([C@H]3[C@@H]([C@]4(C)C(=CC3)C[C@@H](O)CC4)CC2)CC1 ORNBQBCIOKFOEO-YQUGOWONSA-N 0.000 description 3
- 102100031463 Serine/threonine-protein kinase PLK1 Human genes 0.000 description 3
- 108020004459 Small interfering RNA Proteins 0.000 description 3
- 102100029152 UDP-glucuronosyltransferase 1A1 Human genes 0.000 description 3
- 101710205316 UDP-glucuronosyltransferase 1A1 Proteins 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 210000002459 blastocyst Anatomy 0.000 description 3
- 125000004093 cyano group Chemical group *C#N 0.000 description 3
- 125000000753 cycloalkyl group Chemical group 0.000 description 3
- 238000013537 high throughput screening Methods 0.000 description 3
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 210000005228 liver tissue Anatomy 0.000 description 3
- 238000012423 maintenance Methods 0.000 description 3
- 108010056274 polo-like kinase 1 Proteins 0.000 description 3
- 239000002243 precursor Substances 0.000 description 3
- 229960000249 pregnenolone Drugs 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 125000006274 (C1-C3)alkoxy group Chemical group 0.000 description 2
- FPNZBYLXNYPRLR-UHFFFAOYSA-N 2-(4-carbamimidoylphenyl)-1h-indole-6-carboximidamide;hydron;dichloride Chemical compound Cl.Cl.C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FPNZBYLXNYPRLR-UHFFFAOYSA-N 0.000 description 2
- SUBDBMMJDZJVOS-UHFFFAOYSA-N 5-methoxy-2-{[(4-methoxy-3,5-dimethylpyridin-2-yl)methyl]sulfinyl}-1H-benzimidazole Chemical compound N=1C2=CC(OC)=CC=C2NC=1S(=O)CC1=NC=C(C)C(OC)=C1C SUBDBMMJDZJVOS-UHFFFAOYSA-N 0.000 description 2
- 101001059929 Caenorhabditis elegans Forkhead box protein O Proteins 0.000 description 2
- 108010075016 Ceruloplasmin Proteins 0.000 description 2
- 102100023321 Ceruloplasmin Human genes 0.000 description 2
- 208000005038 Cytochrome P-450 CYP1A2 Inducers Diseases 0.000 description 2
- 108010020070 Cytochrome P-450 CYP2B6 Proteins 0.000 description 2
- 102000009666 Cytochrome P-450 CYP2B6 Human genes 0.000 description 2
- 101710176143 Cytochrome P450 7A1 Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 108010049003 Fibrinogen Proteins 0.000 description 2
- 102000008946 Fibrinogen Human genes 0.000 description 2
- 108090000852 Forkhead Transcription Factors Proteins 0.000 description 2
- 102000004315 Forkhead Transcription Factors Human genes 0.000 description 2
- 108010086524 Hepatocyte Nuclear Factor 4 Proteins 0.000 description 2
- 101000957672 Homo sapiens Cytochrome P450 7A1 Proteins 0.000 description 2
- 101000691614 Homo sapiens Serine/threonine-protein kinase PLK3 Proteins 0.000 description 2
- 101000582914 Homo sapiens Serine/threonine-protein kinase PLK4 Proteins 0.000 description 2
- 108090001030 Lipoproteins Proteins 0.000 description 2
- 102000004895 Lipoproteins Human genes 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 102100037223 Nuclear receptor coactivator 1 Human genes 0.000 description 2
- 102100030569 Nuclear receptor corepressor 2 Human genes 0.000 description 2
- 101710153660 Nuclear receptor corepressor 2 Proteins 0.000 description 2
- 102100028960 Peroxisome proliferator-activated receptor gamma coactivator 1-alpha Human genes 0.000 description 2
- 241000288906 Primates Species 0.000 description 2
- 102100026209 Serine/threonine-protein kinase PLK3 Human genes 0.000 description 2
- 102100030267 Serine/threonine-protein kinase PLK4 Human genes 0.000 description 2
- 108091027967 Small hairpin RNA Proteins 0.000 description 2
- 102000007451 Steroid Receptors Human genes 0.000 description 2
- 108010085012 Steroid Receptors Proteins 0.000 description 2
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 2
- 102000004338 Transferrin Human genes 0.000 description 2
- 108090000901 Transferrin Proteins 0.000 description 2
- 239000012190 activator Substances 0.000 description 2
- 125000003545 alkoxy group Chemical group 0.000 description 2
- 125000000033 alkoxyamino group Chemical group 0.000 description 2
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 2
- 239000007640 basal medium Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000003855 cell nucleus Anatomy 0.000 description 2
- 230000036755 cellular response Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 230000003081 coactivator Effects 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 229940012952 fibrinogen Drugs 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 230000030279 gene silencing Effects 0.000 description 2
- 108091008634 hepatocyte nuclear factors 4 Proteins 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- 238000012744 immunostaining Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000006698 induction Effects 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
- 108010082117 matrigel Proteins 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 210000002894 multi-fate stem cell Anatomy 0.000 description 2
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 229960000381 omeprazole Drugs 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 230000003389 potentiating effect Effects 0.000 description 2
- ORNBQBCIOKFOEO-QGVNFLHTSA-N pregnenolone Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 ORNBQBCIOKFOEO-QGVNFLHTSA-N 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 230000008672 reprogramming Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 102000003702 retinoic acid receptors Human genes 0.000 description 2
- 108090000064 retinoic acid receptors Proteins 0.000 description 2
- 239000004055 small Interfering RNA Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 125000001424 substituent group Chemical group 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 125000005505 thiomorpholino group Chemical group 0.000 description 2
- 102000004217 thyroid hormone receptors Human genes 0.000 description 2
- 108090000721 thyroid hormone receptors Proteins 0.000 description 2
- 231100000041 toxicology testing Toxicity 0.000 description 2
- 239000012581 transferrin Substances 0.000 description 2
- 125000000876 trifluoromethoxy group Chemical group FC(F)(F)O* 0.000 description 2
- 125000002023 trifluoromethyl group Chemical group FC(F)(F)* 0.000 description 2
- 230000035899 viability Effects 0.000 description 2
- 229930003231 vitamin Natural products 0.000 description 2
- 239000011782 vitamin Substances 0.000 description 2
- 229940088594 vitamin Drugs 0.000 description 2
- 235000013343 vitamin Nutrition 0.000 description 2
- 102100028162 ATP-binding cassette sub-family C member 3 Human genes 0.000 description 1
- 102100028163 ATP-binding cassette sub-family C member 4 Human genes 0.000 description 1
- 102100033350 ATP-dependent translocase ABCB1 Human genes 0.000 description 1
- 102000001714 Agammaglobulinaemia Tyrosine Kinase Human genes 0.000 description 1
- 108010029445 Agammaglobulinaemia Tyrosine Kinase Proteins 0.000 description 1
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 1
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 1
- 102000005369 Aldehyde Dehydrogenase Human genes 0.000 description 1
- 108020002663 Aldehyde Dehydrogenase Proteins 0.000 description 1
- 108700023418 Amidases Proteins 0.000 description 1
- 108010032178 Amino-acid N-acetyltransferase Proteins 0.000 description 1
- 102000007610 Amino-acid N-acetyltransferase Human genes 0.000 description 1
- 101000993093 Arabidopsis thaliana Heat stress transcription factor B-2a Proteins 0.000 description 1
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 1
- 102100028282 Bile salt export pump Human genes 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 101150051438 CYP gene Proteins 0.000 description 1
- 102000007132 Carboxyl and Carbamoyl Transferases Human genes 0.000 description 1
- 108010072957 Carboxyl and Carbamoyl Transferases Proteins 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010074918 Cytochrome P-450 CYP1A1 Proteins 0.000 description 1
- 108010026925 Cytochrome P-450 CYP2C19 Proteins 0.000 description 1
- 108010000561 Cytochrome P-450 CYP2C8 Proteins 0.000 description 1
- 108010000543 Cytochrome P-450 CYP2C9 Proteins 0.000 description 1
- 208000000130 Cytochrome P-450 CYP3A Inducers Diseases 0.000 description 1
- 102100031476 Cytochrome P450 1A1 Human genes 0.000 description 1
- 102100036194 Cytochrome P450 2A6 Human genes 0.000 description 1
- 102100029363 Cytochrome P450 2C19 Human genes 0.000 description 1
- 102100029359 Cytochrome P450 2C8 Human genes 0.000 description 1
- 102100029358 Cytochrome P450 2C9 Human genes 0.000 description 1
- 102100038696 Cytochrome P450 3A43 Human genes 0.000 description 1
- 102100039203 Cytochrome P450 3A7 Human genes 0.000 description 1
- 102100024918 Cytochrome P450 4F12 Human genes 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 208000030453 Drug-Related Side Effects and Adverse reaction Diseases 0.000 description 1
- 239000006145 Eagle's minimal essential medium Substances 0.000 description 1
- 108020002908 Epoxide hydrolase Proteins 0.000 description 1
- 102000005486 Epoxide hydrolase Human genes 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 108010009306 Forkhead Box Protein O1 Proteins 0.000 description 1
- 108010009307 Forkhead Box Protein O3 Proteins 0.000 description 1
- 102100035427 Forkhead box protein O1 Human genes 0.000 description 1
- 102100035421 Forkhead box protein O3 Human genes 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102100037478 Glutathione S-transferase A2 Human genes 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 102000018997 Growth Hormone Human genes 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- 102100032606 Heat shock factor protein 1 Human genes 0.000 description 1
- 101710190344 Heat shock factor protein 1 Proteins 0.000 description 1
- 206010019695 Hepatic neoplasm Diseases 0.000 description 1
- 102000003745 Hepatocyte Growth Factor Human genes 0.000 description 1
- 108090000100 Hepatocyte Growth Factor Proteins 0.000 description 1
- 102100029283 Hepatocyte nuclear factor 3-alpha Human genes 0.000 description 1
- 102100021374 Hepatocyte nuclear factor 3-gamma Human genes 0.000 description 1
- 101150094793 Hes3 gene Proteins 0.000 description 1
- 101150029234 Hes5 gene Proteins 0.000 description 1
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- 101000986633 Homo sapiens ATP-binding cassette sub-family C member 3 Proteins 0.000 description 1
- 101000986629 Homo sapiens ATP-binding cassette sub-family C member 4 Proteins 0.000 description 1
- 101000875170 Homo sapiens Cytochrome P450 2A6 Proteins 0.000 description 1
- 101000957698 Homo sapiens Cytochrome P450 3A43 Proteins 0.000 description 1
- 101000745715 Homo sapiens Cytochrome P450 3A7 Proteins 0.000 description 1
- 101000909108 Homo sapiens Cytochrome P450 4F12 Proteins 0.000 description 1
- 101001026115 Homo sapiens Glutathione S-transferase A2 Proteins 0.000 description 1
- 101001062353 Homo sapiens Hepatocyte nuclear factor 3-alpha Proteins 0.000 description 1
- 101000818741 Homo sapiens Hepatocyte nuclear factor 3-gamma Proteins 0.000 description 1
- 101000969812 Homo sapiens Multidrug resistance-associated protein 1 Proteins 0.000 description 1
- 101000602926 Homo sapiens Nuclear receptor coactivator 1 Proteins 0.000 description 1
- 101000974356 Homo sapiens Nuclear receptor coactivator 3 Proteins 0.000 description 1
- 101001123331 Homo sapiens Peroxisome proliferator-activated receptor gamma coactivator 1-alpha Proteins 0.000 description 1
- 101000955481 Homo sapiens Phosphatidylcholine translocator ABCB4 Proteins 0.000 description 1
- 101000651467 Homo sapiens Proto-oncogene tyrosine-protein kinase Src Proteins 0.000 description 1
- 101000826399 Homo sapiens Sulfotransferase 1A1 Proteins 0.000 description 1
- 101000826397 Homo sapiens Sulfotransferase 1A2 Proteins 0.000 description 1
- 101000826408 Homo sapiens Sulfotransferase 1B1 Proteins 0.000 description 1
- 101000585344 Homo sapiens Sulfotransferase 1E1 Proteins 0.000 description 1
- 101000585365 Homo sapiens Sulfotransferase 2A1 Proteins 0.000 description 1
- 101000843556 Homo sapiens Transcription factor HES-1 Proteins 0.000 description 1
- 101000912503 Homo sapiens Tyrosine-protein kinase Fgr Proteins 0.000 description 1
- 101000956263 Homo sapiens Uncharacterized protein C19orf48 Proteins 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 108010047230 Member 1 Subfamily B ATP Binding Cassette Transporter Proteins 0.000 description 1
- 108010093662 Member 11 Subfamily B ATP Binding Cassette Transporter Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000010909 Monoamine Oxidase Human genes 0.000 description 1
- 108010062431 Monoamine oxidase Proteins 0.000 description 1
- 102100021339 Multidrug resistance-associated protein 1 Human genes 0.000 description 1
- 101710202061 N-acetyltransferase Proteins 0.000 description 1
- 108010045510 NADPH-Ferrihemoprotein Reductase Proteins 0.000 description 1
- 102100023897 NADPH-cytochrome P450 reductase Human genes 0.000 description 1
- 101710198130 NADPH-cytochrome P450 reductase Proteins 0.000 description 1
- 108090001146 Nuclear Receptor Coactivator 1 Proteins 0.000 description 1
- 102100037226 Nuclear receptor coactivator 2 Human genes 0.000 description 1
- 102100022883 Nuclear receptor coactivator 3 Human genes 0.000 description 1
- 108010016731 PPAR gamma Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 108090000310 Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha Proteins 0.000 description 1
- 102000003728 Peroxisome Proliferator-Activated Receptors Human genes 0.000 description 1
- 108090000029 Peroxisome Proliferator-Activated Receptors Proteins 0.000 description 1
- 102100038825 Peroxisome proliferator-activated receptor gamma Human genes 0.000 description 1
- 102100039032 Phosphatidylcholine translocator ABCB4 Human genes 0.000 description 1
- 101150104557 Ppargc1a gene Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 108090000315 Protein Kinase C Proteins 0.000 description 1
- 102000003923 Protein Kinase C Human genes 0.000 description 1
- 102000055027 Protein Methyltransferases Human genes 0.000 description 1
- 108700040121 Protein Methyltransferases Proteins 0.000 description 1
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 1
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 101100016889 Rattus norvegicus Hes2 gene Proteins 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 238000011579 SCID mouse model Methods 0.000 description 1
- 108091006296 SLC2A1 Proteins 0.000 description 1
- 108091006731 SLCO1B1 Proteins 0.000 description 1
- 108091006686 SLCO2B1 Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102100023536 Solute carrier family 2, facilitated glucose transporter member 1 Human genes 0.000 description 1
- 102100027233 Solute carrier organic anion transporter family member 1B1 Human genes 0.000 description 1
- 102100027264 Solute carrier organic anion transporter family member 2B1 Human genes 0.000 description 1
- 208000007271 Substance Withdrawal Syndrome Diseases 0.000 description 1
- 102100023986 Sulfotransferase 1A1 Human genes 0.000 description 1
- 102100023984 Sulfotransferase 1A2 Human genes 0.000 description 1
- 102100023983 Sulfotransferase 1A3 Human genes 0.000 description 1
- 102100023988 Sulfotransferase 1B1 Human genes 0.000 description 1
- 102100029862 Sulfotransferase 1E1 Human genes 0.000 description 1
- 102100029867 Sulfotransferase 2A1 Human genes 0.000 description 1
- 108010017842 Telomerase Proteins 0.000 description 1
- 206010043276 Teratoma Diseases 0.000 description 1
- 206010070863 Toxicity to various agents Diseases 0.000 description 1
- 102100030798 Transcription factor HES-1 Human genes 0.000 description 1
- 102100040198 UDP-glucuronosyltransferase 1-6 Human genes 0.000 description 1
- 102100040210 UDP-glucuronosyltransferase 1A8 Human genes 0.000 description 1
- 102100029633 UDP-glucuronosyltransferase 2B15 Human genes 0.000 description 1
- 101710200683 UDP-glucuronosyltransferase 2B15 Proteins 0.000 description 1
- 102100029785 UDP-glucuronosyltransferase 2B4 Human genes 0.000 description 1
- 101710200334 UDP-glucuronosyltransferase 2B4 Proteins 0.000 description 1
- 101710008381 UGT1A6 Proteins 0.000 description 1
- 108010074998 UGT1A8 UDP-glucuronosyltransferase Proteins 0.000 description 1
- 102100038573 Uncharacterized protein C19orf48 Human genes 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 101001062354 Xenopus tropicalis Forkhead box protein A2 Proteins 0.000 description 1
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 102000005922 amidase Human genes 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 210000004271 bone marrow stromal cell Anatomy 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 238000003763 carbonization Methods 0.000 description 1
- 238000012832 cell culture technique Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000002875 cyclin dependent kinase inhibitor Substances 0.000 description 1
- 229940043378 cyclin-dependent kinase inhibitor Drugs 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 210000003981 ectoderm Anatomy 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 210000001900 endoderm Anatomy 0.000 description 1
- 239000003623 enhancer Substances 0.000 description 1
- 230000001973 epigenetic effect Effects 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000004129 fatty acid metabolism Effects 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000008571 general function Effects 0.000 description 1
- 210000001654 germ layer Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000014101 glucose homeostasis Effects 0.000 description 1
- 230000004153 glucose metabolism Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 208000006359 hepatoblastoma Diseases 0.000 description 1
- 229940121372 histone deacetylase inhibitor Drugs 0.000 description 1
- 239000003276 histone deacetylase inhibitor Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 150000004677 hydrates Chemical class 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229910001410 inorganic ion Inorganic materials 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 108010093564 inter-alpha-inhibitor Proteins 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000031142 liver development Effects 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 1
- 210000003716 mesoderm Anatomy 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 108010083982 monoamine-sulfating phenol sulfotransferase Proteins 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 230000001323 posttranslational effect Effects 0.000 description 1
- 125000003461 pregnenolone group Chemical group 0.000 description 1
- 239000013630 prepared media Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 229940043437 protein kinase A inhibitor Drugs 0.000 description 1
- 150000003212 purines Chemical class 0.000 description 1
- 150000003230 pyrimidines Chemical class 0.000 description 1
- 230000022983 regulation of cell cycle Effects 0.000 description 1
- 230000009711 regulatory function Effects 0.000 description 1
- 150000004492 retinoid derivatives Chemical class 0.000 description 1
- 102000027483 retinoid hormone receptors Human genes 0.000 description 1
- 108091008679 retinoid hormone receptors Proteins 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 239000012453 solvate Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 210000003014 totipotent stem cell Anatomy 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 230000006648 viral gene expression Effects 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
Images
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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5067—Liver cells
-
- 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
-
- 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
- C12N2503/00—Use of cells in diagnostics
- C12N2503/02—Drug screening
-
- 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
- C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
- C12N2506/02—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from embryonic cells
-
- 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
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/50—Proteins
- C12N2533/54—Collagen; Gelatin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/902—Oxidoreductases (1.)
- G01N2333/90245—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- General Engineering & Computer Science (AREA)
- Hematology (AREA)
- Physics & Mathematics (AREA)
- Urology & Nephrology (AREA)
- Biophysics (AREA)
- Tropical Medicine & Parasitology (AREA)
- Toxicology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Description
R1は、(i)(C1〜C3)アルキル、(ii)(C1〜C3)シクロアルキル、(iii)ハロ、ニトロ、シアノ、ヒドロキシル、トリフルオロメチル、トリフルオロメトキシ、(C1〜C3)アルコキシ、(C1〜C3)アルキル、(C1〜C3)アルカノイル、−S(O)2Rc、もしくはNRaRbから独立して選択される1つ以上の置換基により任意選択で置換されたフェニル、または(iv)NRaRbであり;
R2は、ヒドロキシル、(C1〜C6)アルコキシ、(C1〜C6)アルカノキシアミノ(alkanoxy amino)(C2〜C5)アルコキシ;ヒドロキシ(C2〜C5)アルコキシアミノ(C2〜C5)アルカノキシ、またはヒドロキシル(C2〜C5)アルカノキシであり;
R3は、シアノまたは(C1〜C3)アルカノイルであり;
R4は、水素、(C1〜C3)アルキル、ヒドロキシル(C2〜C5)アルキル、またはアミノ(C2〜C5)アルキルであり;
RaおよびRbは、それぞれ、独立して、水素もしくは(C1〜C3)アルキルであるか、またはRaおよびRbは、それらが結合した窒素が加わって、ピロリジノ、ピペリジノ、モルホリノ、もしくはチオモルホリノであり;
nおよびmは、独立して、0〜2の整数であり;
Xは、ハロであり;
Yは、−S(O)2Rcまたは−ORcであり;
Rcは、(i)非置換もしくはハロにより置換された(C1〜C3)アルキルまたは(ii)アリールである)
による化合物がBTK活性を有することが期待されるであろう。
nおよびmは、独立して、0〜2の整数であり、
Xは、ハロであり、
Yは、−S(O)2Rcまたは−ORcであり、
Rcは、(i)非置換もしくはハロにより置換された(C1〜C3)アルキルまたは(ii)アリールである)
による化合物を含む。
R1は、(i)(C1〜C3)アルキル、(ii)(C1〜C3)シクロアルキル、(iii)ハロ、ニトロ、シアノ、ヒドロキシル、トリフルオロメチル、トリフルオロメトキシ、(C1〜C3)アルコキシ、(C1〜C3)アルキル、(C1〜C3)アルカノイル、−S(O)2Rc、もしくはNRaRbから独立して選択される1つ以上の置換基により任意選択で置換されたフェニル、または(iv)NRaRbであり;
R2は、ヒドロキシル、(C1〜C6)アルコキシ、(C1〜C6)アルカノキシアミノ(C2〜C5)アルコキシ;ヒドロキシ(C2〜C5)アルコキシアミノ(C2〜C5)アルカノキシ、またはヒドロキシル(C2〜C5)アルカノキシであり;
R3は、シアノまたは(C1〜C3)アルカノイルであり;
R4は、水素、(C1〜C3)アルキル、ヒドロキシル(C2〜C5)アルキル、またはアミノ(C2〜C5)アルキルであり;
RaおよびRbは、それぞれ、独立して、水素もしくは(C1〜C3)アルキルであるか、またはRaおよびRbは、それらが結合した窒素が加わって、ピロリジノ、ピペリジノ、モルホリノ、もしくはチオモルホリノであり;
nおよびmは、独立して、0〜2の整数であり;
Xは、ハロであり;
Yは、−S(O)2Rcまたは−ORcであり;
Rcは、(i)非置換もしくはハロにより置換された(C1〜C3)アルキルまたは(ii)アリールである)
の構造を有する化合物を含む。組成物はまた、細胞のためのエネルギー源、アミノ酸、ビタミン、またはその他同種のものなどの細胞培養の1つ以上の構成成分を含むことができる。化合物は、培養肝臓細胞の代謝機能を増強するのに十分な量で組成物中に存在することができる。
nおよびmは、独立して、0〜2の整数であり、
Xは、ハロであり、
Yは、−S(O)2Rcまたは−ORcであり、
Rcは、(i)非置換もしくはハロにより置換された(C1〜C3)アルキルまたは(ii)アリールである)
の構造を有する、第1の態様による組成物組成物である。
初代ヒト肝細胞におけるCYP3A4およびCYP1A2の機能に対するレフルノミド(LEF)、レフルノミド代謝物質(LFM)、およびレフルノミド代謝物質アナログA−13(LFMA−13)の効果を試験する。手短に言えば、コラーゲンIコート表面上で培養した初代ヒト肝細胞を、それぞれ、ビヒクルコントロール、10μM LF、10μM LFM、および10μM LFM−A13を含有するCorning肝細胞培地とインキュベートする。48時間のインキュベーションの終わりに、CYP3A4およびCYP1A2酵素アッセイをPromega P450−Glo(商標)アッセイキットを使用して実行する。アッセイ結果は、同じウェルにおいてATPレベルに対して標準化する。結果を図1に示す。
ヒト胚性幹細胞(hESC)に由来する肝細胞に対するLFM−A13の効果について評価する。手短に言えば、hESCを、当業者に知られている方法を使用して肝細胞系列にコミットさせ、次いで、10μM LFM−A13の存在下または非存在下においてCorning肝細胞培地中で培養する。7日後に、幹細胞に由来する肝細胞は、肝細胞タンパク質の発現を検出するために;(b)肝臓前駆体マーカーであるアルブミン(ALB)の存在について試験するために;ならびに(c)ATP結合カセット(ABC)トランスポーターのスーパーファミリーのメンバーおよび肝機能についてマーカーである多剤耐性関連タンパク質2(MRP2)の存在について試験するために、一次抗体により免疫染色する。細胞の核は、すべての細胞を視覚化するために4’,6−ジアミジノ−2−フェニルインドールジヒドロクロリド(DAPI)により対比染色する(図a)。免疫染色の結果を図9に示す。示されるように、LFM−A13の非存在下において培養されたhESC(左の列)は、低レベルの肝臓マーカー(ALBおよびMRP2)を有する。対照的に、LFM−A13の存在下において培養されたhESC(右の列)は、より高いレベルの肝臓マーカー(ALBおよびMRP2)を有する。LFM−A13が肝細胞へのhESCの分化、維持、または機能を容易にしたことを示す。
Claims (12)
- 肝臓細胞を培養するための組成物において、
肝臓細胞と、
LFM−A13と、
を含むことを特徴とする組成物。 - 前記LFM−A13が、0.1マイクロモル〜50マイクロモルの濃度で前記組成物中に存在することを特徴とする、請求項1に記載の組成物。
- 前記LFM−A13が、1マイクロモル〜20マイクロモルの濃度で前記組成物中に存在することを特徴とする、請求項1に記載の組成物。
- 前記組成物が血清を含むことを特徴とする、請求項1に記載の組成物。
- 前記組成物が血清を含まないことを特徴とする、請求項1に記載の組成物。
- 前記肝臓細胞が肝細胞であることを特徴とする、請求項1〜5のいずれか一項に記載の組成物。
- 前記肝臓細胞が肝細胞に由来する不死化細胞を含むことを特徴とする、請求項1〜5のいずれか一項に記載の組成物。
- 前記肝臓細胞が細胞株を含むことを特徴とする、請求項1〜5のいずれか一項に記載の組成物。
- 前記細胞株が肝細胞に由来することを特徴とする、請求項8に記載の組成物。
- 候補化合物であって、その代謝物質が分析されることが所望される候補化合物をさらに含むことを特徴とする、請求項1〜9のいずれか一項に記載の組成物。
- 培養中の肝臓細胞の代謝機能を増強する方法において、
請求項1〜9のいずれか一項に記載の組成物中で肝臓細胞を培養するステップであって、前記LFM−A13は、実質的に類似するが、LFM−A13を欠く培地中で培養された肝臓細胞に比べて、前記培養肝臓細胞における2つ以上の薬物代謝酵素のレベルを増強するのに有効な量で存在するステップ、
を有してなることを特徴とする方法。 - 候補化合物の代謝物質を同定する方法において、
請求項1〜9のいずれか一項に記載の組成物中で肝臓細胞を培養するステップであって、前記LFM−A13は、実質的に類似するが、LFM−A13を欠く組成物中で培養された肝臓細胞に比べて、前記培養肝臓細胞における2つ以上の薬物代謝酵素のレベルを増強するのに有効な量で存在するステップ、
前記培養細胞を前記候補化合物と接触させるステップ、および
前記候補化合物の代謝物質を同定するステップ、
を有してなることを特徴とする方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201261730762P | 2012-11-28 | 2012-11-28 | |
US61/730,762 | 2012-11-28 | ||
PCT/US2013/071636 WO2014085308A1 (en) | 2012-11-28 | 2013-11-25 | Cell culture medium for enhanced hepatocyte function |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2015536656A JP2015536656A (ja) | 2015-12-24 |
JP6307515B2 true JP6307515B2 (ja) | 2018-04-04 |
Family
ID=49724697
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2015544169A Active JP6307515B2 (ja) | 2012-11-28 | 2013-11-25 | 肝細胞機能の増強のための細胞培養培地 |
Country Status (5)
Country | Link |
---|---|
US (1) | US10876096B2 (ja) |
EP (1) | EP2925858B1 (ja) |
JP (1) | JP6307515B2 (ja) |
CN (1) | CN105189737B (ja) |
WO (1) | WO2014085308A1 (ja) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016167953A1 (en) | 2015-04-16 | 2016-10-20 | Biotranex, Llc | Method of measuring inhibition of phosphatidylcholine export transport and/or formation activity |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20010042804A (ko) * | 1998-04-17 | 2001-05-25 | 파커 휴우즈 인스티튜트 | 비티케이 억제제와 그들의 동정 방법 및 용도 |
US6303652B1 (en) * | 1998-08-21 | 2001-10-16 | Hughes Institute | BTK inhibitors and methods for their identification and use |
AU2003228348A1 (en) | 2002-03-20 | 2003-10-08 | John T. Fassett | Induction of hepatocyte proliferation in vitro by inhibition of cell cycle inhibitors |
WO2005033294A2 (en) * | 2003-09-30 | 2005-04-14 | Regents Of The University Of California | Methods for maintaining hepatocytes in culture and for differentiating embryonic stem cells along a hepatocyte lineage |
CA2562343A1 (en) * | 2004-04-07 | 2005-10-27 | Gene Logic, Inc. | Hepatotoxicity molecular models |
PL2516626T3 (pl) * | 2009-12-23 | 2017-10-31 | Janssen Biotech Inc | Różnicowanie ludzkich zarodkowych komórek macierzystych |
-
2013
- 2013-11-25 CN CN201380071463.0A patent/CN105189737B/zh active Active
- 2013-11-25 JP JP2015544169A patent/JP6307515B2/ja active Active
- 2013-11-25 EP EP13801961.7A patent/EP2925858B1/en active Active
- 2013-11-25 WO PCT/US2013/071636 patent/WO2014085308A1/en active Application Filing
- 2013-11-25 US US14/647,304 patent/US10876096B2/en active Active
Also Published As
Publication number | Publication date |
---|---|
US20150291932A1 (en) | 2015-10-15 |
CN105189737A (zh) | 2015-12-23 |
US10876096B2 (en) | 2020-12-29 |
JP2015536656A (ja) | 2015-12-24 |
EP2925858B1 (en) | 2019-05-15 |
EP2925858A1 (en) | 2015-10-07 |
CN105189737B (zh) | 2019-02-12 |
WO2014085308A1 (en) | 2014-06-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Afshari et al. | Different approaches for transformation of mesenchymal stem cells into hepatocyte-like cells | |
Chen et al. | Biotechnology challenges to in vitro maturation of hepatic stem cells | |
US10106777B2 (en) | Directed differentiation and maturation of pluripotent cells into hepatocyte like cells by modulation of Wnt-signalling pathway | |
Sullivan et al. | Generation of functional human hepatic endoderm from human induced pluripotent stem cells | |
Gage et al. | Initial cell seeding density influences pancreatic endocrine development during in vitro differentiation of human embryonic stem cells | |
Lee et al. | Generation of uniform liver spheroids from human pluripotent stem cells for imaging-based drug toxicity analysis | |
Kondo et al. | Histone deacetylase inhibitor valproic acid promotes the differentiation of human induced pluripotent stem cells into hepatocyte-like cells | |
van Wenum et al. | Oxygen drives hepatocyte differentiation and phenotype stability in liver cell lines | |
Roelandt et al. | Directed differentiation of pluripotent stem cells to functional hepatocytes | |
Kimura et al. | Small molecule AT7867 proliferates PDX1-expressing pancreatic progenitor cells derived from human pluripotent stem cells | |
Tang et al. | Chemical cocktails enable hepatic reprogramming of human urine-derived cells with a single transcription factor | |
EP3365429B1 (en) | Production of virus-receptive pluripotent stem cell (psc)-derived hepatocytes | |
Yang et al. | Stage specific reprogramming of mouse embryo liver cells to a beta cell-like phenotype | |
Kang et al. | Hepatic population derived from human pluripotent stem cells is effectively increased by selective removal of undifferentiated stem cells using YM155 | |
Raggi et al. | Leveraging interacting signaling pathways to robustly improve the quality and yield of human pluripotent stem cell-derived hepatoblasts and hepatocytes | |
Olsavsky Goyak et al. | Hepatocyte differentiation | |
Gómez-Lechón et al. | Evaluation of cytochrome P450 activities in human hepatocytes in vitro | |
JP6307515B2 (ja) | 肝細胞機能の増強のための細胞培養培地 | |
WO2003080649A2 (en) | Induction of hepatocyte proliferation in vitro by inhibition of cell cycle inhibitors | |
Sone et al. | Recovery of mature hepatocytic phenotype following bile ductular transdifferentiation of rat hepatocytes in vitro | |
Kaserman et al. | Protocol for directed differentiation of human induced pluripotent stem cells (iPSCs) to a hepatic lineage | |
Blyszczuk et al. | In vitro differentiation of embryonic stem cells into the pancreatic lineage | |
JP2020533025A (ja) | 肝疾患および機能不全を処置するための組成物および方法 | |
Luo et al. | Functional hit 1 (FH1)-based rapid and efficient generation of functional hepatocytes from human mesenchymal stem cells: a novel strategy for hepatic differentiation | |
Radaszkiewicz et al. | The acceleration of cardiomyogenesis in embryonic stem cells in vitro by serum depletion does not increase the number of developed cardiomyocytes |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20160920 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20170718 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20171018 |
|
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: 20180213 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20180312 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 6307515 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313113 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |