WO2017110724A1 - 血液細胞を用いた組織・臓器の作製方法 - Google Patents
血液細胞を用いた組織・臓器の作製方法 Download PDFInfo
- Publication number
- WO2017110724A1 WO2017110724A1 PCT/JP2016/087739 JP2016087739W WO2017110724A1 WO 2017110724 A1 WO2017110724 A1 WO 2017110724A1 JP 2016087739 W JP2016087739 W JP 2016087739W WO 2017110724 A1 WO2017110724 A1 WO 2017110724A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cells
- organ
- tissue
- bud
- cell
- Prior art date
Links
- 210000000056 organ Anatomy 0.000 title claims abstract description 167
- 238000000034 method Methods 0.000 title claims abstract description 82
- 210000000601 blood cell Anatomy 0.000 title claims abstract description 45
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 23
- 210000004027 cell Anatomy 0.000 claims abstract description 192
- 210000001519 tissue Anatomy 0.000 claims abstract description 76
- 241001465754 Metazoa Species 0.000 claims abstract description 34
- 210000003556 vascular endothelial cell Anatomy 0.000 claims abstract description 32
- 238000000338 in vitro Methods 0.000 claims abstract description 24
- 239000003814 drug Substances 0.000 claims abstract description 20
- 238000012258 culturing Methods 0.000 claims abstract description 14
- 229940079593 drug Drugs 0.000 claims abstract description 14
- 230000006870 function Effects 0.000 claims abstract description 14
- 210000003958 hematopoietic stem cell Anatomy 0.000 claims description 33
- 210000004185 liver Anatomy 0.000 claims description 26
- 210000003494 hepatocyte Anatomy 0.000 claims description 19
- 239000003550 marker Substances 0.000 claims description 17
- 108090000623 proteins and genes Proteins 0.000 claims description 15
- 230000004069 differentiation Effects 0.000 claims description 13
- 210000001778 pluripotent stem cell Anatomy 0.000 claims description 13
- 108010088751 Albumins Proteins 0.000 claims description 12
- 102000009027 Albumins Human genes 0.000 claims description 12
- 210000003999 epithelial cell of bile duct Anatomy 0.000 claims description 9
- 210000004700 fetal blood Anatomy 0.000 claims description 9
- 230000014509 gene expression Effects 0.000 claims description 8
- 102100023635 Alpha-fetoprotein Human genes 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 7
- 210000004263 induced pluripotent stem cell Anatomy 0.000 claims description 5
- 230000028327 secretion Effects 0.000 claims description 5
- 102100039203 Cytochrome P450 3A7 Human genes 0.000 claims description 4
- 101000745715 Homo sapiens Cytochrome P450 3A7 Proteins 0.000 claims description 4
- 101000892398 Homo sapiens Tryptophan 2,3-dioxygenase Proteins 0.000 claims description 4
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims description 4
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims description 4
- 102100040653 Tryptophan 2,3-dioxygenase Human genes 0.000 claims description 4
- 108010026331 alpha-Fetoproteins Proteins 0.000 claims description 4
- 238000011084 recovery Methods 0.000 claims description 4
- 230000033667 organ regeneration Effects 0.000 claims description 3
- 230000017423 tissue regeneration Effects 0.000 claims description 3
- 210000005087 mononuclear cell Anatomy 0.000 claims description 2
- 108010003524 sodium-bile acid cotransporter Proteins 0.000 claims description 2
- 230000001172 regenerating effect Effects 0.000 abstract description 5
- 230000009467 reduction Effects 0.000 abstract description 3
- 241000282472 Canis lupus familiaris Species 0.000 description 12
- 241000251730 Chondrichthyes Species 0.000 description 12
- 241000238557 Decapoda Species 0.000 description 11
- 230000035800 maturation Effects 0.000 description 11
- 210000000130 stem cell Anatomy 0.000 description 11
- 241000282412 Homo Species 0.000 description 10
- 238000011156 evaluation Methods 0.000 description 10
- 230000002440 hepatic effect Effects 0.000 description 10
- 210000001616 monocyte Anatomy 0.000 description 10
- 210000004369 blood Anatomy 0.000 description 9
- 239000008280 blood Substances 0.000 description 9
- 238000005516 engineering process Methods 0.000 description 9
- 241000699670 Mus sp. Species 0.000 description 8
- 210000004039 endoderm cell Anatomy 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- -1 AFP Proteins 0.000 description 6
- 241000972773 Aulopiformes Species 0.000 description 6
- 241000283690 Bos taurus Species 0.000 description 6
- 241000282693 Cercopithecidae Species 0.000 description 6
- 241000777300 Congiopodidae Species 0.000 description 6
- 241000283086 Equidae Species 0.000 description 6
- 241000287828 Gallus gallus Species 0.000 description 6
- 102100035716 Glycophorin-A Human genes 0.000 description 6
- 241000283973 Oryctolagus cuniculus Species 0.000 description 6
- 241001494479 Pecora Species 0.000 description 6
- 241000700159 Rattus Species 0.000 description 6
- 241000282887 Suidae Species 0.000 description 6
- 238000010171 animal model Methods 0.000 description 6
- 210000004556 brain Anatomy 0.000 description 6
- 235000013330 chicken meat Nutrition 0.000 description 6
- 210000001900 endoderm Anatomy 0.000 description 6
- 230000000968 intestinal effect Effects 0.000 description 6
- 210000003734 kidney Anatomy 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 235000019515 salmon Nutrition 0.000 description 6
- 210000000278 spinal cord Anatomy 0.000 description 6
- 230000002792 vascular Effects 0.000 description 6
- 238000004113 cell culture Methods 0.000 description 5
- 230000036267 drug metabolism Effects 0.000 description 5
- 210000001035 gastrointestinal tract Anatomy 0.000 description 5
- 210000002216 heart Anatomy 0.000 description 5
- 210000003887 myelocyte Anatomy 0.000 description 5
- 239000002243 precursor Substances 0.000 description 5
- 238000012216 screening Methods 0.000 description 5
- 210000003954 umbilical cord Anatomy 0.000 description 5
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- 108091005250 Glycophorins Proteins 0.000 description 4
- 101000711846 Homo sapiens Transcription factor SOX-9 Proteins 0.000 description 4
- 102100034204 Transcription factor SOX-9 Human genes 0.000 description 4
- 210000001185 bone marrow Anatomy 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 210000001671 embryonic stem cell Anatomy 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 4
- 239000012139 lysis buffer Substances 0.000 description 4
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 4
- 210000000496 pancreas Anatomy 0.000 description 4
- 210000000952 spleen Anatomy 0.000 description 4
- 238000002054 transplantation Methods 0.000 description 4
- 206010013530 Diverticula Diseases 0.000 description 3
- 206010013554 Diverticulum Diseases 0.000 description 3
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 3
- 206010019663 Hepatic failure Diseases 0.000 description 3
- 101500025419 Homo sapiens Epidermal growth factor Proteins 0.000 description 3
- 101001078143 Homo sapiens Integrin alpha-IIb Proteins 0.000 description 3
- 101000800116 Homo sapiens Thy-1 membrane glycoprotein Proteins 0.000 description 3
- 102100025306 Integrin alpha-IIb Human genes 0.000 description 3
- 108090000630 Oncostatin M Proteins 0.000 description 3
- 102100033523 Thy-1 membrane glycoprotein Human genes 0.000 description 3
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 3
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 3
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 3
- 102100033177 Vascular endothelial growth factor receptor 2 Human genes 0.000 description 3
- 210000003651 basophil Anatomy 0.000 description 3
- 230000000747 cardiac effect Effects 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000007876 drug discovery Methods 0.000 description 3
- 210000002889 endothelial cell Anatomy 0.000 description 3
- 230000002327 eosinophilic effect Effects 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- 210000002149 gonad Anatomy 0.000 description 3
- 229940116978 human epidermal growth factor Drugs 0.000 description 3
- 208000007903 liver failure Diseases 0.000 description 3
- 231100000835 liver failure Toxicity 0.000 description 3
- 210000004072 lung Anatomy 0.000 description 3
- 108010082117 matrigel Proteins 0.000 description 3
- GVUGOAYIVIDWIO-UFWWTJHBSA-N nepidermin Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CCC(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](CC=1C=CC(O)=CC=1)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](CCCCN)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)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CS)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CS)NC(=O)[C@H](C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)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=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C(C)C)C(C)C)C1=CC=C(O)C=C1 GVUGOAYIVIDWIO-UFWWTJHBSA-N 0.000 description 3
- 210000002220 organoid Anatomy 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- 210000001685 thyroid gland Anatomy 0.000 description 3
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 102100031585 ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 Human genes 0.000 description 2
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 2
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 2
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 2
- 108050005493 CD3 protein, epsilon/gamma/delta subunit Proteins 0.000 description 2
- 108010083123 CDX2 Transcription Factor Proteins 0.000 description 2
- 102100025470 Carcinoembryonic antigen-related cell adhesion molecule 8 Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 206010013710 Drug interaction Diseases 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 description 2
- 102100035961 Hematopoietically-expressed homeobox protein HHEX Human genes 0.000 description 2
- 102100022054 Hepatocyte nuclear factor 4-alpha Human genes 0.000 description 2
- 102100031671 Homeobox protein CDX-2 Human genes 0.000 description 2
- 101000777636 Homo sapiens ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 Proteins 0.000 description 2
- 101000693913 Homo sapiens Albumin Proteins 0.000 description 2
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 2
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 2
- 101000914320 Homo sapiens Carcinoembryonic antigen-related cell adhesion molecule 8 Proteins 0.000 description 2
- 101001074244 Homo sapiens Glycophorin-A Proteins 0.000 description 2
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 description 2
- 101001021503 Homo sapiens Hematopoietically-expressed homeobox protein HHEX Proteins 0.000 description 2
- 101001045740 Homo sapiens Hepatocyte nuclear factor 4-alpha Proteins 0.000 description 2
- 101000994365 Homo sapiens Integrin alpha-6 Proteins 0.000 description 2
- 101000608935 Homo sapiens Leukosialin Proteins 0.000 description 2
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 description 2
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 description 2
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 2
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 description 2
- 101001070790 Homo sapiens Platelet glycoprotein Ib alpha chain Proteins 0.000 description 2
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 2
- 101000884271 Homo sapiens Signal transducer CD24 Proteins 0.000 description 2
- 101000633780 Homo sapiens Signaling lymphocytic activation molecule Proteins 0.000 description 2
- 101000914496 Homo sapiens T-cell antigen CD7 Proteins 0.000 description 2
- 101000934346 Homo sapiens T-cell surface antigen CD2 Proteins 0.000 description 2
- 101000716102 Homo sapiens T-cell surface glycoprotein CD4 Proteins 0.000 description 2
- 101000946843 Homo sapiens T-cell surface glycoprotein CD8 alpha chain Proteins 0.000 description 2
- 101000652324 Homo sapiens Transcription factor SOX-17 Proteins 0.000 description 2
- 101000687905 Homo sapiens Transcription factor SOX-2 Proteins 0.000 description 2
- 101000835093 Homo sapiens Transferrin receptor protein 1 Proteins 0.000 description 2
- 101000851007 Homo sapiens Vascular endothelial growth factor receptor 2 Proteins 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 102100032816 Integrin alpha-6 Human genes 0.000 description 2
- 102100039564 Leukosialin Human genes 0.000 description 2
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 description 2
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 102000003729 Neprilysin Human genes 0.000 description 2
- 108090000028 Neprilysin Proteins 0.000 description 2
- 108010088225 Nestin Proteins 0.000 description 2
- 102000008730 Nestin Human genes 0.000 description 2
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 2
- 102100041030 Pancreas/duodenum homeobox protein 1 Human genes 0.000 description 2
- 102100034173 Platelet glycoprotein Ib alpha chain Human genes 0.000 description 2
- 102000016971 Proto-Oncogene Proteins c-kit Human genes 0.000 description 2
- 108010014608 Proto-Oncogene Proteins c-kit Proteins 0.000 description 2
- 101710183548 Pyridoxal 5'-phosphate synthase subunit PdxS Proteins 0.000 description 2
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 2
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 2
- 102100038081 Signal transducer CD24 Human genes 0.000 description 2
- 102100029215 Signaling lymphocytic activation molecule Human genes 0.000 description 2
- 102100027208 T-cell antigen CD7 Human genes 0.000 description 2
- 102100025237 T-cell surface antigen CD2 Human genes 0.000 description 2
- 102100036011 T-cell surface glycoprotein CD4 Human genes 0.000 description 2
- 102100034922 T-cell surface glycoprotein CD8 alpha chain Human genes 0.000 description 2
- 102100030243 Transcription factor SOX-17 Human genes 0.000 description 2
- 102100024270 Transcription factor SOX-2 Human genes 0.000 description 2
- 102100026144 Transferrin receptor protein 1 Human genes 0.000 description 2
- 229960005070 ascorbic acid Drugs 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 210000002808 connective tissue Anatomy 0.000 description 2
- 210000003890 endocrine cell Anatomy 0.000 description 2
- 230000003511 endothelial effect Effects 0.000 description 2
- 210000003762 eosinophilic myelocyte Anatomy 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 210000003238 esophagus Anatomy 0.000 description 2
- 230000029142 excretion Effects 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 102000044814 human ALB Human genes 0.000 description 2
- 229960000890 hydrocortisone Drugs 0.000 description 2
- 230000001939 inductive effect 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
- 238000002955 isolation Methods 0.000 description 2
- 210000003593 megakaryocyte Anatomy 0.000 description 2
- 210000000713 mesentery Anatomy 0.000 description 2
- 210000003716 mesoderm Anatomy 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 210000001167 myeloblast Anatomy 0.000 description 2
- 210000005055 nestin Anatomy 0.000 description 2
- 210000004498 neuroglial cell Anatomy 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 210000003924 normoblast Anatomy 0.000 description 2
- 230000000849 parathyroid Effects 0.000 description 2
- 230000010412 perfusion Effects 0.000 description 2
- 210000003240 portal vein Anatomy 0.000 description 2
- 230000008569 process Effects 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
- 238000011160 research Methods 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- WBWWGRHZICKQGZ-HZAMXZRMSA-N taurocholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCS(O)(=O)=O)C)[C@@]2(C)[C@@H](O)C1 WBWWGRHZICKQGZ-HZAMXZRMSA-N 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 210000001541 thymus gland Anatomy 0.000 description 2
- 210000003606 umbilical vein Anatomy 0.000 description 2
- BJHCYTJNPVGSBZ-YXSASFKJSA-N 1-[4-[6-amino-5-[(Z)-methoxyiminomethyl]pyrimidin-4-yl]oxy-2-chlorophenyl]-3-ethylurea Chemical compound CCNC(=O)Nc1ccc(Oc2ncnc(N)c2\C=N/OC)cc1Cl BJHCYTJNPVGSBZ-YXSASFKJSA-N 0.000 description 1
- DFFCDIMSXJYMTB-AATRIKPKSA-N 2-[(e)-2-(6-carbamimidoyl-1h-benzimidazol-2-yl)ethenyl]-3h-benzimidazole-5-carboximidamide Chemical compound C1=C(C(N)=N)C=C2NC(/C=C/C3=NC4=CC=C(C=C4N3)C(=N)N)=NC2=C1 DFFCDIMSXJYMTB-AATRIKPKSA-N 0.000 description 1
- ZDTNHRWWURISAA-UHFFFAOYSA-N 4',5'-dibromo-3',6'-dihydroxyspiro[2-benzofuran-3,9'-xanthene]-1-one Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C(Br)=C1OC1=C(Br)C(O)=CC=C21 ZDTNHRWWURISAA-UHFFFAOYSA-N 0.000 description 1
- 102100022464 5'-nucleotidase Human genes 0.000 description 1
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 1
- 108010059616 Activins Proteins 0.000 description 1
- 102100032964 Alpha-actinin-2 Human genes 0.000 description 1
- APKFDSVGJQXUKY-KKGHZKTASA-N Amphotericin-B Natural products O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1C=CC=CC=CC=CC=CC=CC=C[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-KKGHZKTASA-N 0.000 description 1
- 102100022014 Angiopoietin-1 receptor Human genes 0.000 description 1
- 102000007372 Ataxin-1 Human genes 0.000 description 1
- 108010032963 Ataxin-1 Proteins 0.000 description 1
- 102100024505 Bone morphogenetic protein 4 Human genes 0.000 description 1
- 102100031650 C-X-C chemokine receptor type 4 Human genes 0.000 description 1
- 102100032912 CD44 antigen Human genes 0.000 description 1
- 102000000905 Cadherin Human genes 0.000 description 1
- 108050007957 Cadherin Proteins 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 108020003264 Cotransporters Proteins 0.000 description 1
- 102000034534 Cotransporters Human genes 0.000 description 1
- 102100037241 Endoglin Human genes 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 1
- 102100037362 Fibronectin Human genes 0.000 description 1
- 108010067306 Fibronectins Proteins 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- 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 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 1
- 102100021866 Hepatocyte growth factor Human genes 0.000 description 1
- 101150068639 Hnf4a gene Proteins 0.000 description 1
- 102100028404 Homeobox protein Hox-B4 Human genes 0.000 description 1
- 102100027875 Homeobox protein Nkx-2.5 Human genes 0.000 description 1
- 102100027332 Homeobox protein SIX2 Human genes 0.000 description 1
- 101000678236 Homo sapiens 5'-nucleotidase Proteins 0.000 description 1
- 101000797275 Homo sapiens Alpha-actinin-2 Proteins 0.000 description 1
- 101000753291 Homo sapiens Angiopoietin-1 receptor Proteins 0.000 description 1
- 101000762379 Homo sapiens Bone morphogenetic protein 4 Proteins 0.000 description 1
- 101000922348 Homo sapiens C-X-C chemokine receptor type 4 Proteins 0.000 description 1
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 1
- 101000881679 Homo sapiens Endoglin Proteins 0.000 description 1
- 101000898034 Homo sapiens Hepatocyte growth factor Proteins 0.000 description 1
- 101000839788 Homo sapiens Homeobox protein Hox-B4 Proteins 0.000 description 1
- 101000632197 Homo sapiens Homeobox protein Nkx-2.5 Proteins 0.000 description 1
- 101000651912 Homo sapiens Homeobox protein SIX2 Proteins 0.000 description 1
- 101000599951 Homo sapiens Insulin-like growth factor I Proteins 0.000 description 1
- 101000935043 Homo sapiens Integrin beta-1 Proteins 0.000 description 1
- 101001076408 Homo sapiens Interleukin-6 Proteins 0.000 description 1
- 101001120813 Homo sapiens Myosin regulatory light chain 2, atrial isoform Proteins 0.000 description 1
- 101000958741 Homo sapiens Myosin-6 Proteins 0.000 description 1
- 101000610551 Homo sapiens Prominin-1 Proteins 0.000 description 1
- 101000740205 Homo sapiens Sal-like protein 1 Proteins 0.000 description 1
- 101000868152 Homo sapiens Son of sevenless homolog 1 Proteins 0.000 description 1
- 101000801254 Homo sapiens Tumor necrosis factor receptor superfamily member 16 Proteins 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- 102100026818 Inhibin beta E chain Human genes 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 102100037852 Insulin-like growth factor I Human genes 0.000 description 1
- 102100025304 Integrin beta-1 Human genes 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- 108010085895 Laminin Proteins 0.000 description 1
- 102000007547 Laminin Human genes 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 102100026057 Myosin regulatory light chain 2, atrial isoform Human genes 0.000 description 1
- 102100038319 Myosin-6 Human genes 0.000 description 1
- 102100037369 Nidogen-1 Human genes 0.000 description 1
- 102000004140 Oncostatin M Human genes 0.000 description 1
- BELBBZDIHDAJOR-UHFFFAOYSA-N Phenolsulfonephthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 BELBBZDIHDAJOR-UHFFFAOYSA-N 0.000 description 1
- 102100024616 Platelet endothelial cell adhesion molecule Human genes 0.000 description 1
- 102100040120 Prominin-1 Human genes 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 108091006611 SLC10A1 Proteins 0.000 description 1
- 102100037204 Sal-like protein 1 Human genes 0.000 description 1
- 102100021988 Sodium/bile acid cotransporter Human genes 0.000 description 1
- 240000006394 Sorghum bicolor Species 0.000 description 1
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 1
- 208000009415 Spinocerebellar Ataxias Diseases 0.000 description 1
- WBWWGRHZICKQGZ-UHFFFAOYSA-N Taurocholic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(=O)NCCS(O)(=O)=O)C)C1(C)C(O)C2 WBWWGRHZICKQGZ-UHFFFAOYSA-N 0.000 description 1
- 208000026062 Tissue disease Diseases 0.000 description 1
- 102100033725 Tumor necrosis factor receptor superfamily member 16 Human genes 0.000 description 1
- 108010053096 Vascular Endothelial Growth Factor Receptor-1 Proteins 0.000 description 1
- 108010053099 Vascular Endothelial Growth Factor Receptor-2 Proteins 0.000 description 1
- 108010053100 Vascular Endothelial Growth Factor Receptor-3 Proteins 0.000 description 1
- 102100033178 Vascular endothelial growth factor receptor 1 Human genes 0.000 description 1
- 102100033179 Vascular endothelial growth factor receptor 3 Human genes 0.000 description 1
- 102100035140 Vitronectin Human genes 0.000 description 1
- 108010031318 Vitronectin Proteins 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000000488 activin Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 210000000577 adipose tissue Anatomy 0.000 description 1
- 210000002556 adrenal cortex cell Anatomy 0.000 description 1
- 210000001943 adrenal medulla Anatomy 0.000 description 1
- 210000001132 alveolar macrophage Anatomy 0.000 description 1
- APKFDSVGJQXUKY-INPOYWNPSA-N amphotericin B Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/C=C/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-INPOYWNPSA-N 0.000 description 1
- 229960003942 amphotericin b Drugs 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 210000001608 connective tissue cell Anatomy 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 210000000695 crystalline len Anatomy 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 210000003074 dental pulp Anatomy 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 210000004207 dermis Anatomy 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 238000009513 drug distribution Methods 0.000 description 1
- 210000003981 ectoderm Anatomy 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 210000003989 endothelium vascular Anatomy 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 210000003237 epithelioid cell Anatomy 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 210000001647 gastrula Anatomy 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 230000001434 glomerular Effects 0.000 description 1
- 229960002743 glutamine Drugs 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 210000004209 hair Anatomy 0.000 description 1
- 210000003566 hemangioblast Anatomy 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 229920006130 high-performance polyamide Polymers 0.000 description 1
- 210000003630 histaminocyte Anatomy 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 1
- 238000007917 intracranial administration Methods 0.000 description 1
- 210000001865 kupffer cell Anatomy 0.000 description 1
- 210000001821 langerhans cell Anatomy 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000005033 mesothelial cell Anatomy 0.000 description 1
- 230000007102 metabolic function Effects 0.000 description 1
- 210000004925 microvascular endothelial cell Anatomy 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 210000000885 nephron Anatomy 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 108010008217 nidogen Proteins 0.000 description 1
- 239000012038 nucleophile Substances 0.000 description 1
- 210000002997 osteoclast Anatomy 0.000 description 1
- 210000000277 pancreatic duct Anatomy 0.000 description 1
- 230000009996 pancreatic endocrine effect Effects 0.000 description 1
- 210000002990 parathyroid gland Anatomy 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 210000000578 peripheral nerve Anatomy 0.000 description 1
- 229960003531 phenolsulfonphthalein Drugs 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 210000005059 placental tissue Anatomy 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 210000004765 promyelocyte Anatomy 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 210000001995 reticulocyte Anatomy 0.000 description 1
- 210000004116 schwann cell Anatomy 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 210000000697 sensory organ Anatomy 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 210000002356 skeleton Anatomy 0.000 description 1
- 210000003625 skull Anatomy 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 201000003624 spinocerebellar ataxia type 1 Diseases 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
- 239000003106 tissue adhesive Substances 0.000 description 1
- 229940075469 tissue adhesives Drugs 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000004926 tubular epithelial cell Anatomy 0.000 description 1
- 210000000626 ureter Anatomy 0.000 description 1
- 210000001635 urinary tract Anatomy 0.000 description 1
- 230000000982 vasogenic effect 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
- C12N5/0697—Artificial constructs associating cells of different lineages, e.g. tissue equivalents
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0271—Chimeric vertebrates, e.g. comprising exogenous cells
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
-
- 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
- C12N5/0062—General methods for three-dimensional culture
-
- 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
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0647—Haematopoietic stem cells; Uncommitted or multipotent progenitors
-
- 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
- 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
- C12N5/0676—Pancreatic cells
- C12N5/0677—Three-dimensional culture, tissue culture or organ culture; Encapsulated 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
- 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
- C12N5/069—Vascular Endothelial 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
- 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
- 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
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
-
- 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
- G01N33/5082—Supracellular entities, e.g. tissue, organisms
- G01N33/5088—Supracellular entities, e.g. tissue, organisms of vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/12—Animals modified by administration of exogenous cells
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/02—Animal zootechnically ameliorated
- A01K2267/025—Animal producing cells or organs for transplantation
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
-
- 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
- C12N2502/00—Coculture with; Conditioned medium produced by
- C12N2502/11—Coculture with; Conditioned medium produced by blood or immune system 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
- C12N2502/00—Coculture with; Conditioned medium produced by
- C12N2502/11—Coculture with; Conditioned medium produced by blood or immune system cells
- C12N2502/1171—Haematopoietic stem cells
Definitions
- the present invention relates to a tissue / organ preparation method using blood cells.
- Non-patent Document 1 Takebe T. et.al., Nature (2013), Patent Document 1: Human tissue / organ production method WO2013 / 047639 A1).
- Non-Patent Document 2 Kamiya A., et al., EMBO J. 1999 18: 2127-3
- Non-Patent Document 3 Kinoshita T., et al., Proc Natl Acad Sci U S A. 1999 96: 7265-70 (1999)).
- the present invention aims to provide such an improved technique.
- the present inventors succeeded in further improving the function of the formed organ bud by adding hematopoietic cells to a culture system in which hepatic endoderm cells, vascular endothelial cells and mesenchymal cells are co-cultured.
- hematopoietic cells a culture system in which hepatic endoderm cells, vascular endothelial cells and mesenchymal cells are co-cultured.
- the gist of the present invention is as follows.
- the blood cells comprise undifferentiated hematopoietic cells.
- the undifferentiated hematopoietic cells are hematopoietic progenitor cells and / or hematopoietic stem cells.
- the blood cells are derived from umbilical cord blood.
- the blood cells are cells of a mononuclear cell fraction of cord blood.
- the tissue or organ cell is an undifferentiated cell derived from a pluripotent stem cell.
- the pluripotent stem cell is an induced pluripotent stem cell.
- the pluripotent stem cell is derived from human.
- (13) Compared with hepatic buds prepared by culturing vascular endothelial cells, mesenchymal cells and tissue or organ cells in vitro in the absence of blood cells, The method according to (12), which is prepared.
- hepatocyte differentiation marker gene Increased expression of hepatocyte differentiation marker gene in the absence of blood cells compared to hepatoblasts prepared by culturing vascular endothelial cells, mesenchymal cells and tissue or organ cells in vitro
- the hepatocyte differentiation marker gene is at least one marker selected from the group consisting of ⁇ -fetoprotein, albumin, CYP3A7, tryptophan metabolizing enzyme TDO2, and sodium taurocholate cotransporter.
- An organ bud produced by the method according to any one of (1) to (15).
- a method for producing a tissue or organ comprising transplanting the organ bud according to (16) into a non-human animal and allowing it to differentiate into a tissue or organ.
- a method for transplanting an organ bud comprising transplanting the organ bud according to (16) to a human or non-human animal.
- a tissue or organ regeneration or function recovery method comprising transplanting the organ bud according to (16) into a human or non-human animal and differentiating it into a tissue or organ.
- a method for producing a non-human chimeric animal comprising transplanting the organ bud according to (16) into a non-human animal and allowing it to differentiate into a tissue or an organ.
- (21) at least one selected from the group consisting of organ buds according to (16), tissues and organs produced by the method according to (17), and non-human chimeric animals produced by the method according to (20) How to use and evaluate drugs.
- Non-Patent Document 2 Kamiya A). Et al., EMBO J. 1999 18: 2127-3
- Non-Patent Document 3 Kinoshita T., et al., Proc Natl Acad Sci U S A. 1999 96: 7265-70 (1999)).
- OSM cytokines
- Patent Document 1 Human tissue / organ production method WO2013 / 047639 A1
- Patent Document 2 Human tissue / organ production method WO2013 / 047639 A1
- the present invention provides a method for producing organ buds, comprising culturing vascular endothelial cells, mesenchymal cells, and tissue or organ cells in vitro in the presence of blood cells.
- an “organ bud” is a structure that can be differentiated into an organ upon maturation, and is a tissue or organ cell, vascular endothelial cell, mesenchymal cell (undifferentiated mesenchymal cell or differentiated therefrom) Cell) and a structure containing four types of blood cells. Whether a structure is an organ bud is examined, for example, by examining whether the structure can be transplanted into a living body and differentiated into the target organ (if it has differentiated into the target organ, it is determined to be an organ bud) And / or whether or not the structure includes all the four types of cells described above (if all the four types of cells are included, it can be determined that the bud is an organ bud).
- Organ buds may be organ buds that differentiate into organs such as kidney, heart, lung, spleen, esophagus, stomach, thyroid, parathyroid, thymus, gonad, brain, spinal cord, etc., but differentiate into liver Organ buds that differentiate into endoderm organs such as organ buds (liver buds), organ buds that differentiate into pancreas (pancreatic buds), and organ buds that differentiate into the intestinal tract are preferred. Whether or not a certain structure is an organ bud that differentiates into an endoderm organ can be confirmed by examining the expression of a marker protein (if one or more of the marker proteins described below are expressed, the organ bud Can be determined.) For example, in liver buds, HHEX, SOX2, HNF4A, AFP, ALB, etc.
- liverlivebud liver diverticula, liver organoid, pancreatic (dorsal or ventral) buds, pancreatic diverticula, pancreatic organoid, intestinal bud, intestinal diverticula, intestinal organoid (K. Matsumoto, et al .Science.19; 294 (5542): 559-63. (2001)) and the like are included in the organ buds of the present invention.
- the “blood cell” is a cell isolated from the living body, a cell induced to differentiate from a stem cell such as an ES cell or iPS cell, or a cell directly reprogrammed by gene transfer into the differentiated cell. Having blood cell characteristics (eg, CD34, CD2, CD3, CD4, CD7, CD8, CD10, CD14, CD16, CD19, CD20, CD24, CD41, CD45, CD56, CD66b, or CD235a, CD38, CD90, CD49f , VEGFR2, CD43, CD71, GPA (Glycophorin A), CD42b, c-kit, CD150, Sca-1, Ter119, etc. cells).
- a stem cell such as an ES cell or iPS cell
- a cell is a blood cell is CD34, CD2, CD3, CD4, CD7, CD8, CD10, CD14, CD16, CD19, CD20, CD24, CD41, CD45, CD56, CD66b, or CD235a, CD38, CD90 , CD49f, VEGFR2, CD43, CD71, GPA (Glycophorin A), CD42b, c-kit, CD150, Sca-1, Ter119, etc.
- the blood cells used in the present invention may be differentiated or undifferentiated, but preferably contain undifferentiated hematopoietic cells such as hematopoietic progenitor cells and hematopoietic stem cells.
- Undifferentiated hematopoietic cells are collected from pluripotent stem cells such as induced pluripotent stem cells (iPS cells) and embryonic stem cells (ES cells), and blood (umbilical cord blood, bone marrow blood, peripheral blood, etc.) And blood cells that have been directly transdifferentiated (directly reprogrammed) from other differentiated cells.
- pluripotent stem cells such as induced pluripotent stem cells (iPS cells) and embryonic stem cells (ES cells)
- iPS cells induced pluripotent stem cells
- ES cells embryonic stem cells
- blood umbilical cord blood, bone marrow blood, peripheral blood, etc.
- blood cells that have been directly transdifferentiated (directly reprogrammed) from other differentiated cells In the Examples described later, it is considered that hematopoietic progenitor cells and hematopoietic stem cells could be concentrated by taking a monocyte fraction of cord blood.
- hematopoietic stem cells blood stem cells, hematopoietic progenitor cells, blood progenitor cells, myeloid progenitor cells, granulocyte / monocyte progenitor cells, granulocyte progenitor cells, granulocytes, bone marrow Blast, promyelocyte, myelocyte, retromyelocyte, rod-shaped nucleus cell, segmented nucleus cell (neutrophil), monocyte progenitor cell, monocyte, macrophage, histocyte, Kupffer cell, alveolar macrophage, small Glial cells, osteoclasts, epithelioid cells, giant cells (Langhans giant cells, foreign body giant cells, Tohton giant cells), dendritic cells, Langerhans cells, myeloid monocytes, myeloblasts, basophilic promyelo Sphere, basophil myelocyte, basophil postmyelocyte, basophil,
- Blood cells are mainly derived from humans, but animals other than humans (for example, animals used for laboratory animals, pets, working animals, racehorses, dogs, etc., specifically mice, rats, rabbits) , Pigs, dogs, monkeys, cows, horses, sheep, chickens, sharks, rays, sharks, salmon, shrimps, crabs, etc.).
- vascular endothelial cell refers to a cell constituting the vascular endothelium or a cell that can differentiate into such a cell. Whether a cell is a vascular endothelial cell can be confirmed by examining whether a marker protein, for example, TIE2, VEGFR-1, VEGFR-2, VEGFR-3, or CD41 is expressed (any of the above-mentioned marker proteins). If one or more of them are expressed, it can be judged as a vascular endothelial cell).
- vascular endothelial cells used in the present invention may be differentiated or undifferentiated. Whether vascular endothelial cells are differentiated cells can be confirmed by CD31 and CD144.
- endothelial cells endothelial cells, umbilical vein endothelial cells, endothelial progenitor cells, endothelial precursor cells, vasculogenic progenitors, hemangioblast (HJ. Joo, et al. Blood. 25; 4 8: 8; -104. (2011)) is included in the vascular endothelial cells in the present invention.
- Preferred vascular endothelial cells are umbilical vein-derived vascular endothelial cells.
- Vascular endothelial cells can be collected from blood vessels or prepared from pluripotent stem cells such as induced pluripotent stem cells (iPS cells) and embryonic stem cells (ES cells) according to known methods.
- Vascular endothelial cells are mainly derived from humans, but animals other than humans (for example, animals used for experimental animals, pets, working animals, racehorses, dogs, etc., specifically mice, rats, Rabbits, pigs, dogs, monkeys, cows, horses, sheep, chickens, sharks, rays, sharks, salmons, shrimps, crabs, etc.) may be used.
- animals other than humans for example, animals used for experimental animals, pets, working animals, racehorses, dogs, etc., specifically mice, rats, Rabbits, pigs, dogs, monkeys, cows, horses, sheep, chickens, sharks, rays, sharks, salmons, shrimps, crabs, etc.
- the term “mesenchymal cell” refers to a connective tissue cell that is present in connective tissue mainly derived from mesoderm and forms a support structure for cells that function in the tissue. Is a concept that includes cells that have not yet differentiated into mesenchymal cells.
- the mesenchymal cells used in the present invention may be differentiated or undifferentiated. Whether a cell is an undifferentiated mesenchymal cell is confirmed by examining whether a marker protein such as Stro-1, CD29, CD44, CD73, CD90, CD105, CD133, CD271, or Nestin is expressed (If any one or more of the marker proteins are expressed, it can be determined that they are undifferentiated mesenchymal cells).
- a mesenchymal cell that does not express any of the markers in the preceding paragraph can be determined as a differentiated mesenchymal cell.
- mesenchymal stem cells mesenchymal progenitor cells, mesenchymal cells (R. Peters, et al. PLoS One. 30; 5 (12): e15689. (2010)) Included in mesenchymal cells.
- Preferred mesenchymal cells are bone marrow-derived mesenchymal cells (particularly mesenchymal stem cells).
- Mesenchymal cells are collected from bone marrow, adipose tissue, placental tissue, umbilical cord tissue, dental pulp, or other pluripotent cells such as artificial pluripotent stem cells (iPS cells) and embryonic stem cells (ES cells). It can be prepared from stem cells according to a known method.
- Mesenchymal cells are mainly derived from humans, but animals other than humans (for example, animals used for laboratory animals, pets, working animals, racehorses, dogs, etc., specifically mice, rats) , Rabbits, pigs, dogs, monkeys, cows, horses, sheep, chickens, sharks, rays, sharks, salmon, shrimps, crabs, etc.).
- tissue or organ cell refers to a functional cell that constitutes a tissue or organ, or an undifferentiated cell that differentiates into a functional cell.
- tissue or organ cells include cells that can differentiate into organs such as kidney, heart, lung, spleen, esophagus, stomach, thyroid, parathyroid, thymus, gonad, brain, spinal cord, etc.
- a cell is a cell that can differentiate into an ectodermal organ, mesodermal organ, or endodermal organ can be confirmed by examining the expression of a marker protein (either one or more of the marker proteins are expressed). If so, it can be determined that the cell can differentiate into an endoderm organ.
- a marker protein either one or more of the marker proteins are expressed.
- HHEX, SOX2, HNF4A, AFP, and ALB are markers
- PDX1, SOX17, SOX9, etc. are markers that can differentiate into the intestinal tract. Then, CDX2, SOX9, etc.
- markers are markers, and in cells that can differentiate into kidney, SIX2, SALL1, in cells that can differentiate into heart, NKX2-5 MYH6, ACTN2, MYL7, HPPA, in cells that can differentiate into blood, For C-KIT, SCA1, TER119, HOXB4, and cells that can differentiate into the brain and spinal cord, HNK1, AP2, NESTIN, etc. are markers.
- hepatoblast hepatic progenitor cells, pancreatoblast, hepatic precursor cells, pancreatoblast, pancreatic progenitors, pancreatic progenitor cells, pancreatic precursor cells, endocrine precursors, intestinal s , Metanephric mesenchymal precursor cells, multipotent nephron progenitor, renal progenitor cell, cardiac mesoderm, cardiovascular progenitor cells, cardiac progenitor cells, (JR. Spence, et al. Nature .; 470 (7332): 105 (elf) , Et al. EMBO J .; 25 (21): 5214-5228. (2006), J.
- Undifferentiated tissue or organ cells are collected from the tissue or organ, or prepared from pluripotent stem cells such as induced pluripotent stem cells (iPS cells) and embryonic stem cells (ES cells) according to known methods. Can do.
- pluripotent stem cells such as induced pluripotent stem cells (iPS cells) and embryonic stem cells (ES cells) according to known methods.
- iPS cells induced pluripotent stem cells
- ES cells embryonic stem cells
- undifferentiated tissue or organ cells are cells that are in the middle of differentiation from pluripotent stem cells such as iPS cells into tissues or organs, such as primitive intestinal endoderm cells (Primitive Gut Endoderm Cells: PGECs) (patents) No. 5777127).
- PGECs can differentiate into hepatocytes, pancreatic cells and intestinal cells (high differentiation function), do not express markers related to malignancy of cancer (high safety), and are non-feeder environment from iPS cells Since it can be prepared by inducing differentiation below, clinical application is also possible.
- PGECs can be prepared in large quantities. PGECs can be prepared by the method described in Japanese Patent No. 5777127.
- both CXCR4 and E-cadherin-positive endoderm cells were induced, and the obtained endoderm It may be a CXCR4-negative HNF4 ⁇ -positive liver endoderm population obtained by adding BMP4 and FGF2 to a cell line and culturing for 2 days.
- organ cells that can differentiate into the liver are K.Si-Taiyeb, et al. Hepatology, 51 (1): 297- 305 (2010), T. Touboul, et al. Hepatology. 51 (5): 1754-65.
- Organ cells that can differentiate into pancreas should be prepared according to D. Zhang, et al. Cell Res.; 19 (4): 429-38. (2009) Organ cells that can differentiate into the intestinal tract are J. Cai, et al. J Mol Cell Biol.; 2 (1): 50-60 (2010), R. Spence, et al. Nature.; 470 (7332 ): 105-9. (2011) and organ cells that can differentiate into the heart can be prepared according to J. Zhang, hanget al. Circulation Research .; 104: e30-e41 (2009) Cells that can differentiate into the brain and spinal cord can be prepared according to G.GLee, et al. Nature Biotechnology 25, 1468-1475 (2007).
- pancreatic endocrine cells pancreatic pancreatic duct epithelial cells
- liver hepatocytes intestinal epithelial cells
- renal tubular epithelial cells renal glomerular epithelial cells
- cardiac myocardial cells examples thereof include blood lymphocytes and granulocytes, red blood cells, brain neurons and glial cells, spinal cord neurons and Schwann cells.
- tissue or organ cells are mainly derived from humans, animals other than humans (for example, animals used for laboratory animals, pets, working animals, racehorses, dogs, etc., specifically mice, rats, etc.) Tissue, organ cells derived from rabbits, pigs, dogs, monkeys, cows, horses, sheep, chickens, sharks, rays, sharks, salmon, shrimps, crabs, etc.).
- Any medium can be used as long as it can form organ buds.
- the medium for vascular endothelial cell culture, the medium for tissue or organ cell culture, and the above two mediums are mixed. It is preferable to use what was made.
- Any medium can be used for vascular endothelial cell culture, but hEGF (recombinant human epidermal growth factor), VEGF (vascular endothelial growth factor), hydrocortisone, bFGF, ascorbic acid, IGF1, FBS Antibiotics (for example, gentamicin, amphotericin B, etc.), Heparin, L-Glutamine, Phenolred, and BBE are preferably used.
- EGM-2 BulletKit (Lonza), EGM BulletKit (Lonza), VascuLife EnGS Comp Kit (LCT), Human Endothelial-SFM Basal Growth Medium (Invitrogen)
- TOYOBO Human microvascular endothelial cell growth medium
- Any medium for tissue or organ cell culture may be used, but when the organ cells are hepatocytes, it contains at least one of ascorbic acid, BSA- FAF, insulin, hydrocortisone, and GA-1000. It is preferable to use one.
- HCM BulletKit (Lonza) minus hEGF (recombinant human epidermal growth factor), RPMI1640 (Sigma-Aldrich) 1% B27 Supplements (GIBCO) And 10 ng / mL HHGF (manufactured by Sigma-Aldrich)
- a mixture of GM BulletKit (Lonza) and HCM BulletKit (Lonza) excluding hEGF (recombinant human epidermal growth factor) in a 1: 1 mixture Addition of Oncostatin M and HGF has been found to be effective in liver bud maturation.
- a scaffold material In the culture of cells, it is not necessary to use a scaffold material, but a mixture of four types of cells may be cultured on a gel-like support capable of contracting mesenchymal cells.
- the contraction of mesenchymal cells is morphologically observed with a microscope or with the naked eye, and shows that the tissue shape is strong enough to maintain the shape of the tissue following recovery with a medicine spoon (Takebe, etc.) et al. Nature 499 (7459), 481-484, 2013)).
- the support has an appropriate hardness (for example, a Young's modulus of 200 kPa or less (in the case of a gel having a flat shape coated with Matrigel, etc.), but the appropriate hardness of the support can vary depending on the coating and shape.)
- the base material is preferably a gel-like base material, and examples of such a base material include hydrogels (eg, acrylamide gel, gelatin, matrigel, etc.), but are not limited thereto. Note that the hardness of the support is not necessarily uniform depending on the shape, size, and quantity of the target assembly, and it is possible to set a spatial or temporal gradient or pattern the hardness. Is possible.
- the hardness of the support is preferably 100 kPa or less, more preferably 1 to 50 kPa.
- the gel-like support may be flat, or the cross-section on the side of the gel-like support to be cultured may be U-shaped or V-shaped. Since the cross-section of the gel-like support on the side to be cultured has a U or V shape, the cells gather on the culture surface of the support, and a cell aggregate is formed with a smaller number of cells and / or tissues. This is advantageous.
- the support may be chemically and physically modified. Examples of the modifying substance include matrigel, laminin, entactin, collagen, fibronectin, vitronectin and the like.
- An example of setting a spatial gradient in the hardness of the gel-like culture support is a gel-like culture support in which the hardness at the center is harder than the hardness at the periphery.
- the center hardness should be 200 kPa or less, and the periphery hardness should be softer than the center.
- the appropriate center and periphery hardness varies depending on the coating and shape. sell.
- Another example in which a spatial gradient is set in the hardness of the gel-like culture support is a gel-like culture support in which the hardness at the periphery is harder than the hardness at the center.
- a patterned gel-like culture support is a gel-like culture support having one or more patterns in which the hardness at the center is harder than the hardness at the periphery.
- the center hardness should be 200 kPa or less, and the periphery hardness should be softer than the center.
- the appropriate center and periphery hardness varies depending on the coating and shape. sell.
- Another example of the patterned gel-like culture support is a gel-type culture support having one or more patterns in which the hardness of the peripheral part is harder than the hardness of the central part.
- the hardness of the periphery is appropriate to be 200 kPa or less, and the hardness of the center should be softer than the periphery.
- the appropriate hardness of the center and periphery of the support varies depending on the coating and shape. sell.
- the temperature during the cultivation is not particularly limited, but is preferably 30 to 40 ° C, more preferably 37 ° C.
- the culture period is not particularly limited, but is preferably 3 to 10 days, and more preferably 6 days.
- Organ buds produced by the method of the present invention have functions compared to organ buds produced by culturing vascular endothelial cells, mesenchymal cells, and tissues or organ cells in vitro in the absence of blood cells. Can be improved.
- the hepatoblast produced by the method of the present invention is obtained by culturing vascular endothelial cells, mesenchymal cells and tissues or organ cells in vitro in the absence of blood cells. Compared with the produced liver bud, the secretory ability of albumin can be improved.
- hepatoblasts produced by the method of the present invention are compared with hepatoblasts produced by culturing vascular endothelial cells, mesenchymal cells and tissues or organ cells in vitro in the absence of blood cells, Expression of hepatocyte differentiation marker gene can be increased.
- hepatocyte differentiation marker genes include, but are not limited to, ⁇ -fetoprotein, albumin, CYP3A7, tryptophan metabolizing enzyme TDO2, sodium / taurocholic acid cotransporter, and the like.
- the present invention also provides an organ bud produced by the above method.
- a tissue or organ can be prepared by transplanting an organ bud prepared by the above method to a non-human animal and allowing it to mature in the non-human animal. That is, the present invention also provides a method for producing a tissue or organ, which comprises transplanting an organ bud produced by the above method into a non-human animal and allowing it to differentiate into a tissue or organ.
- Non-human animals to be used include experimental animals, pets, working animals, racehorses, dogs, etc., specifically mice, rats, rabbits, pigs, dogs, monkeys, cows, horses, sheep , Chicken, shark, ray, shark, salmon, shrimp, crab and the like.
- the non-human animal used is preferably an immunodeficient animal in order to avoid immune rejection.
- the present invention also provides a method for transplanting organ buds, including transplanting the organ buds produced by the above method to a human or non-human animal.
- the transplantation site of the organ bud may be any site as long as transplantation is possible, but examples thereof include intracranial, mesentery, liver, spleen, kidney, subrenal capsule, and portal vein.
- 5mm large organ buds may be transplanted, and when transplanted on the portal vein, about 1 to 20 5mm large organ buds prepared in vitro may be transplanted.
- the tissues and organs produced as described above can be used for drug discovery screening and regenerative medicine.
- the present invention provides a tissue or organ regeneration or function recovery method comprising transplanting an organ bud produced by the above method into a human or non-human animal and differentiating it into a tissue or organ.
- Non-human animals include animals used for laboratory animals, pets, working animals, racehorses, dogs, etc., specifically mice, rats, rabbits, pigs, dogs, monkeys, cows, horses, sheep, chickens. , Sharks, rays, sharks, salmon, shrimps, crabs.
- the organ bud produced by the method of the present invention can be formulated and used in the form of a composition for regenerative medicine.
- the composition of the present invention can be transplanted into a living body to produce a tissue or an organ.
- the composition of the present invention can be transplanted into a living body to regenerate or restore the function of a tissue or organ.
- the organ bud can differentiate into a tissue or organ having a vascular network.
- vascular perfusion can occur in the vascular network.
- tissue / organ having a highly ordered tissue structure equivalent to or close to that of an adult tissue.
- the composition of the present invention includes a tissue vascularization promoter such as FGF2, HGF, and VEGF, a gelatin sponge for hemostasis accompanying transplantation (trade name: Sponzel, Astellas Co., Ltd.), and Bolheel (Teijin) used for fixation of transplanted tissue.
- Tissue adhesives such as Pharma Co., Ltd., Veriplast (CSL Behring Co., Ltd.), and Taco Kombu (CSL Behring Co., Ltd.) may be added.
- the present invention also provides a method for producing a non-human chimeric animal, which comprises transplanting an organ bud produced by the above method into a non-human animal and allowing it to differentiate into a tissue or organ.
- the non-human animal for example, mouse
- transplanted with the organ bud can mimic the physiological function of the tissue species or organ cell-derived species (for example, human) used to produce the organ bud.
- the present invention also provides a method for evaluating a drug using at least one selected from the group consisting of organ buds, tissues and organs produced by the above method, and non-human chimeric animals.
- drug evaluation include evaluation of drug metabolism (for example, prediction of drug metabolism profile), drug efficacy evaluation (for example, screening of drugs effective as pharmaceuticals), toxicity evaluation, drug interaction evaluation, and the like. Can do.
- Drug metabolism is evaluated by collecting a biological sample after administering a drug candidate compound in at least one selected from the group consisting of organ buds, tissues and organs, and non-human chimeric animals prepared by the above method. By analyzing, a human-type drug metabolism profile can be obtained. This makes it possible to predict the distribution, metabolism, and excretion processes of drugs in humans, which was extremely difficult to achieve with conventional technology, and is expected to dramatically accelerate the development of safe and effective drugs.
- Screening for a drug effective as a pharmaceutical is performed by at least one selected from the group consisting of organ buds, tissues and organs, and non-human chimeric animals prepared by the above-described method from cells established from diseased patients. Analysis can be performed by administering the compound. As a result, it is expected that the accuracy of prediction of drug efficacy when administered to humans, which was insufficient in conventional in vitro tests, can be greatly improved.
- Toxicity evaluation is to measure a tissue disorder marker, etc. after administering a test substance to at least one selected from the group consisting of organ buds, tissues and organs prepared by the above method, and non-human chimeric animals. As a result, the accuracy of failure prediction can be improved.
- the drug interaction evaluation is carried out by administering a plurality of drugs using at least one selected from the group consisting of organ buds, tissues and organs, and non-human chimeric animals prepared by the above method, and thereafter It can be performed by evaluating pharmacokinetics such as drug distribution, metabolism, and excretion process, toxicity evaluation, and drug efficacy evaluation.
- tissue stem cells from tissues and organs produced by the method of the present invention, and the present invention can be applied to cell manipulation techniques for mass production of human tissue cells and organ cells.
- Example 1 [Method] 1. Preparation of blood cells ⁇ Acquisition of cord blood> Blood was collected from the umbilical cord (provided by a patient who was delivered by cesarean section at Yokohama City University Hospital) using a 50 mL syringe and 18 G needle. Approximately 40 mL to 50 mL of blood was collected. ⁇ Hemolysis treatment> 40 mL of Lysis buffer was placed in a 50 mL tube, and 10 mL of umbilical cord blood was added. Stir and let stand at room temperature for 10 minutes. Centrifuged at 200G for 10 minutes.
- the suspension was suspended in 10 mL of Lysis buffer to make a total of 50 mL. Again, it was centrifuged at 200G for 10 minutes. The precipitate was suspended in 10 mL DMEM + 10% FBS and centrifuged at 200 G for 5 minutes. Again 10 mL DMEM + 10% FBS was performed and the number of cells was counted.
- PI- / CD235- cells were expanded with FSC (forward scatter) and SSC (side scatter), and the cell population corresponding to the monocyte fraction (MNC) cells was isolated.
- FSC forward scatter
- SSC side scatter
- RPMI + 1% at a cell density of 5x10 4 cells / cm 2 on a plastic dish.
- B27 + 10 uM Rockinhibitor + 50 ng / mL Wnt3a + 100 ng / mL ActivinA was cultured for 1 day. Subsequently, the cells were cultured in the presence of RPMI + 1% B27 + 50 ng / mL Wnt3a + 100 ng / mL ActivinA for 6 days to obtain hepatic endoderm cells.
- [result] 1 ( Figure 1, 2, 3) Hepatic endoderm cells (2.5x10 5 cells) and umbilical cord-derived vascular endothelial cells (1.7x10 5 cells) (Lonza, Basel, Switzerland) and mesenchymal cells (2.5x10 4 cells) (Lonza, Basel, Switzerland) (10: 7: 1) Suspend monocyte fraction (MNC) cells (Fig. 1) containing PI-cells and hematopoietic stem cells / hematopoietic progenitor cells separated using a cell sorter (1x10 5 cells) Liver buds were made on the plate (size: 50-250 micrometers).
- MNC monoocyte fraction
- albumin secretion increased in the group to which monocyte fraction (MNC) cells enriched with PI-cells or hematopoietic stem cells / hematopoietic progenitor cells were added compared to the conventional group without blood cells ( Figure 2).
- MNC monocyte fraction
- hepatic buds on the 15th day of culture cultured under the above conditions were collected, and hepatocyte differentiation markers ( ⁇ -fetoprotein (AFP), albumin (ALB), CYP3A7, tryptophan metabolizing enzyme (TDO2), sodium / taurocholate)
- AFP ⁇ -fetoprotein
- ALB albumin
- CYP3A7 tryptophan metabolizing enzyme
- TDO2 tryptophan metabolizing enzyme
- sodium / taurocholate sodium / taurocholate
- hepatic endoderm cells 2.5 ⁇ 10 5 cells
- umbilical cord-derived endothelial cells 1.7 ⁇ 10 5 cells
- mesenchymal cells 2.5 ⁇ 10 4 cells
- the present invention is an important basic technology for industrial production of human functional cells. Thereby, it can apply to manufacture of the human tissue and organ for transplantation as a regenerative medicine technique for intractable diseases. Not only is it expected to achieve significant cost savings from conventional methods, but it is also possible to improve functional maturity in vitro with this technology, so it was difficult to treat with conventional technology. It is expected to be applicable to acute and subacute liver failure.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Immunology (AREA)
- Hematology (AREA)
- Environmental Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Urology & Nephrology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Animal Husbandry (AREA)
- Animal Behavior & Ethology (AREA)
- Transplantation (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- General Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Pathology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Toxicology (AREA)
- Vascular Medicine (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
(1)血液細胞の存在下で、血管内皮細胞、間葉系細胞、及び組織又は臓器細胞をin vitroで培養することを含む、器官芽の作製方法。
(2)器官芽は、成熟することで器官に分化することができる構造体である(1)記載の方法。
(3)血液細胞の非存在下で、血管内皮細胞、間葉系細胞、及び組織又は臓器細胞をin vitroで培養して作製した器官芽と比較して、機能が向上した器官芽が作製される(1)又は(2)記載の方法。
(4)足場材料を用いることなく、細胞が培養される(1)~(3)のいずれかに記載の方法。
(5)血液細胞が、未分化造血細胞を含む(1)~(4)のいずれかに記載の方法。
(6)未分化造血細胞が、造血前駆細胞及び/又は造血幹細胞である(5)記載の方法。
(7)血液細胞が、臍帯血に由来する(1)~(6)のいずれかに記載の方法。
(8)血液細胞が、臍帯血の単核球画分の細胞である(7)記載の方法。
(9)組織又は臓器細胞が、多能性幹細胞から誘導された未分化細胞である(1)~(8)のいずれかに記載の方法。
(10)多能性幹細胞が人工多能性幹細胞である(9)記載の方法。
(11)多能性幹細胞がヒト由来である(9)又は(10)記載の方法。
(12)器官芽が肝芽である(1)~(11)のいずれかに記載の方法。
(13)血液細胞の非存在下で、血管内皮細胞、間葉系細胞及び組織又は臓器細胞をin vitroで培養して作製した肝芽と比較して、アルブミンの分泌能が向上した肝芽が作製される(12)記載の方法。
(14)血液細胞の非存在下で、血管内皮細胞、間葉系細胞及び組織又は臓器細胞をin vitroで培養して作製した肝芽と比較して、肝細胞分化マーカー遺伝子の発現が上昇した肝芽が作製される(12)又は(13)記載の方法。
(15)肝細胞分化マーカー遺伝子が、αフェトプロテイン、アルブミン、CYP3A7、トリプトファン代謝酵素TDO2、及びナトリウム・タウロコール酸共輸送体からなる群より選択される少なくとも1つのマーカーである(14)記載の方法。
(16)(1)~(15)のいずれかに記載の方法によって作製された器官芽。
(17)(16)記載の器官芽を非ヒト動物に移植し、組織又は臓器に分化させることを含む、組織又は臓器の作製方法。
(18)(16)記載の器官芽をヒト又は非ヒト動物に移植することを含む、器官芽の移植方法。
(19)(16)記載の器官芽をヒト又は非ヒト動物に移植し、組織又は臓器に分化させることを含む、組織又は臓器の再生又は機能回復方法。
(20)(16)記載の器官芽を非ヒト動物に移植し、組織又は臓器に分化させることを含む、非ヒトキメラ動物の作製方法。
(21)(16)記載の器官芽、(17)記載の方法によって作製された組織及び臓器、並びに(20)記載の方法によって作製された非ヒトキメラ動物からなる群より選択される少なくとも一つを用いて、薬剤を評価する方法。
本明細書は、本願の優先権の基礎である日本国特許出願、特願2015‐251140の明細書および/または図面に記載される内容を包含する。
〔実施例1〕
[方法]
1、 血液細胞の調製
<臍帯血の取得>
臍帯(横浜市立大学附属病院にて帝王切開により出産された患者様より提供)より50mLシリンジ、18G針を用いて採血を行った。血液はおおよそ40mL-50mL採取した。
<溶血処理>
50mLチューブに40mLのLysis バッファーを入れ、10mLの臍帯血を加えた。
撹拌し室温で10分間静置した。
200Gで10分遠心分離した。
もう一度、Lysis バッファー10mLに懸濁し、5本分まとめ50mLとした。
再び200Gで10分遠心分離した。
沈殿を10mL DMEM+10%FBSで懸濁し、200Gで5分遠心分離した。
再度10mL DMEM+10%FBSし細胞数を計測した。
材料
1) 10 x Lysis バッファーの作製 (x10 液; 1/10希釈して使用した。)
・NH4Cl 82.6g
・NaHCO3 11.9g
・EDTA2Na 0.378g
/ 1L milliQ adjust to pH7.3
2) 1 x Lysis バッファーの作製
PBS(4℃に冷えている方が良い。2mM EDTAを入れても良い)でx1に希釈して使用した。
<PI-/CD235-細胞の分離>
血液細胞に対しPropidium Iodide (PI)を用いて死細胞を染色し、蛍光標識CD235抗体を用いて赤血球/赤芽球を染色し、セルソーターを用いてそれぞれで染色されない細胞(生細胞で有り、赤血球/赤芽球でない細胞)を分離した。
<monocyte画分(MNC)細胞の分離>
セルソーターを用い、PI-/CD235-細胞をFSC(forward scatter:前方散乱光)、およびSSC(side scatter:側方散乱光)で展開しmonocyte画分(MNC)細胞に該当する細胞集団を分離した。
2、肝内胚葉細胞の作製
方法:
未分化iPS細胞(TkDA3(東京大学より提供)、および、臍帯より独自に樹立したiPS細胞)を剥離し単細胞化した後、プラスチックディッシュに5x104cells/cm2の細胞密度で、RPMI+1%B27+10uM Rockinhibitor+50ng/mL Wnt3a+100ng/mL ActivinA存在下で1日間培養した。続いて、RPMI+1%B27+50ng/mL Wnt3a+ 100ng/mL ActivinA存在下で6日間培養し肝内胚葉細胞を得た。
1(図1、2、3)
肝内胚葉細胞(2.5x105細胞)と臍帯由来血管内皮細胞(1.7x105細胞) (Lonza、Basel、Switzerland)および間葉系細胞(2.5x104細胞) (Lonza、Basel、Switzerland)(10:7:1)に対し、セルソーターを用いて分離したPI-細胞および造血幹細胞/造血前駆細胞が濃縮されている、monocyte画分(MNC)細胞(図1)を1x105細胞懸濁し、クラレマイクロウェルプレート上で肝芽(liver bud)を作製した(大きさ:50 ~250マイクロメートル)。15日間培養後、それぞれの条件で培養した肝芽の培養上清中に24時間で分泌されたヒトアルブミンタンパク質量をELISA法により分析した。その結果PI-細胞あるいは造血幹細胞/造血前駆細胞が濃縮されているmonocyte画分(MNC)細胞を添加した群では、従来法である血液細胞非添加群に較べアルブミン分泌量が増加していた(図2)。次に上記の条件で培養した培養15日目の肝芽を回収しqPCRにより肝細胞分化マーカー(αフェトプロテイン(AFP)、アルブミン(ALB)、CYP3A7、トリプトファン代謝酵素(TDO2)、ナトリウム・タウロコール酸共輸送体(NTCP))の発現を検討したところ、PI-細胞あるいはmonocyte画分(MNC)細胞を添加した群では、従来法である血液細胞非添加群に較べ遺伝子発現量が増加していた(図3)。
2(図4、5)
次に添加する血液細胞の用量依存性を検討するため、肝内胚葉細胞(2.5x105細胞)と臍帯由来血管内皮細胞(1.7x105細胞)および間葉系細胞(2.5x104細胞) (10:7:1)に対し、血液細胞添加ゼロ、血液細胞(PI-細胞)添加1x105細胞、2.5x105細胞、5x105細胞添加群で肝芽作製15日目での培養液(図4)中の24時間ヒトアルブミンタンパク質分泌量を検討した。その結果、従来法に較べ血液細胞の細胞数依存的にヒトアルブミン分泌量が増強されることが明らかとなった(図5)。
本明細書で引用した全ての刊行物、特許および特許出願をそのまま参考として本明細書にとり入れるものとする。
Claims (21)
- 血液細胞の存在下で、血管内皮細胞、間葉系細胞、及び組織又は臓器細胞をin vitroで培養することを含む、器官芽の作製方法。
- 器官芽は、成熟することで器官に分化することができる構造体である請求項1記載の方法。
- 血液細胞の非存在下で、血管内皮細胞、間葉系細胞、及び組織又は臓器細胞をin vitroで培養して作製した器官芽と比較して、機能が向上した器官芽が作製される請求項1又は2記載の方法。
- 足場材料を用いることなく、細胞が培養される請求項1~3のいずれかに記載の方法。
- 血液細胞が、未分化造血細胞を含む請求項1~4のいずれかに記載の方法。
- 未分化造血細胞が、造血前駆細胞及び/又は造血幹細胞である請求項5記載の方法。
- 血液細胞が、臍帯血に由来する請求項1~6のいずれかに記載の方法。
- 血液細胞が、臍帯血の単核球画分の細胞である請求項7記載の方法。
- 組織又は臓器細胞が、多能性幹細胞から誘導された未分化細胞である請求項1~8のいずれかに記載の方法。
- 多能性幹細胞が人工多能性幹細胞である請求項9記載の方法。
- 多能性幹細胞がヒト由来である請求項9又は10記載の方法。
- 器官芽が肝芽である請求項1~11のいずれかに記載の方法。
- 血液細胞の非存在下で、血管内皮細胞、間葉系細胞及び組織又は臓器細胞をin vitroで培養して作製した肝芽と比較して、アルブミンの分泌能が向上した肝芽が作製される請求項12記載の方法。
- 血液細胞の非存在下で、血管内皮細胞、間葉系細胞及び組織又は臓器細胞をin vitroで培養して作製した肝芽と比較して、肝細胞分化マーカー遺伝子の発現が上昇した肝芽が作製される請求項12又は13記載の方法。
- 肝細胞分化マーカー遺伝子が、αフェトプロテイン、アルブミン、CYP3A7、トリプトファン代謝酵素TDO2、及びナトリウム・タウロコール酸共輸送体からなる群より選択される少なくとも1つのマーカーである請求項14記載の方法。
- 請求項1~15のいずれかに記載の方法によって作製された器官芽。
- 請求項16記載の器官芽を非ヒト動物に移植し、組織又は臓器に分化させることを含む、組織又は臓器の作製方法。
- 請求項16記載の器官芽をヒト又は非ヒト動物に移植することを含む、器官芽の移植方法。
- 請求項16記載の器官芽をヒト又は非ヒト動物に移植し、組織又は臓器に分化させることを含む、組織又は臓器の再生又は機能回復方法。
- 請求項16記載の器官芽を非ヒト動物に移植し、組織又は臓器に分化させることを含む、非ヒトキメラ動物の作製方法。
- 請求項16記載の器官芽、請求項17記載の方法によって作製された組織及び臓器、並びに請求項20記載の方法によって作製された非ヒトキメラ動物からなる群より選択される少なくとも一つを用いて、薬剤を評価する方法。
Priority Applications (10)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
BR112018012070-7A BR112018012070A2 (ja) | 2015-12-24 | 2016-12-19 | A manufacturing method of an organization and internal organs using blood cells |
SG11201804755VA SG11201804755VA (en) | 2015-12-24 | 2016-12-19 | Method for manufacturing tissue/organ by using blood cells |
EP16878613.5A EP3395943B1 (en) | 2015-12-24 | 2016-12-19 | Method for manufacturing tissue/organ by using blood cells |
CN201680070326.9A CN108368484B (zh) | 2015-12-24 | 2016-12-19 | 使用了血液细胞的组织、内脏器官的制作方法 |
JP2017558110A JP6860921B2 (ja) | 2015-12-24 | 2016-12-19 | 血液細胞を用いた組織・臓器の作製方法 |
KR1020187017472A KR20180097564A (ko) | 2015-12-24 | 2016-12-19 | 혈액 세포를 사용한 조직·장기의 제조 방법 |
US16/060,814 US20180352792A1 (en) | 2015-12-24 | 2016-12-19 | Method for manufacturing tissue/organ by using blood cells |
AU2016374909A AU2016374909B2 (en) | 2015-12-24 | 2016-12-19 | Method for manufacturing tissue/organ by using blood cells |
CA3008213A CA3008213A1 (en) | 2015-12-24 | 2016-12-19 | Method for manufacturing tissue/organ by using blood cells |
HK18116553.5A HK1257345A1 (zh) | 2015-12-24 | 2018-12-24 | 使用了血液細胞的組織、內臟器官的製作方法 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2015251140 | 2015-12-24 | ||
JP2015-251140 | 2015-12-24 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2017110724A1 true WO2017110724A1 (ja) | 2017-06-29 |
Family
ID=59090367
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2016/087739 WO2017110724A1 (ja) | 2015-12-24 | 2016-12-19 | 血液細胞を用いた組織・臓器の作製方法 |
Country Status (11)
Country | Link |
---|---|
US (1) | US20180352792A1 (ja) |
EP (1) | EP3395943B1 (ja) |
JP (1) | JP6860921B2 (ja) |
KR (1) | KR20180097564A (ja) |
CN (1) | CN108368484B (ja) |
AU (1) | AU2016374909B2 (ja) |
BR (1) | BR112018012070A2 (ja) |
CA (1) | CA3008213A1 (ja) |
HK (1) | HK1257345A1 (ja) |
SG (1) | SG11201804755VA (ja) |
WO (1) | WO2017110724A1 (ja) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2019189324A1 (ja) * | 2018-03-28 | 2019-10-03 | 公立大学法人横浜市立大学 | 細胞塊融合法 |
WO2020175594A1 (ja) * | 2019-02-28 | 2020-09-03 | 公立大学法人横浜市立大学 | 血液凝固および/または補体異常疾患の治療用組成物 |
CN114585746A (zh) * | 2019-10-17 | 2022-06-03 | 公立大学法人横滨市立大学 | 药物毒性评价方法 |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2018181342A1 (ja) * | 2017-03-28 | 2018-10-04 | 味の素株式会社 | 未分化維持培地添加剤 |
EP3949730A4 (en) * | 2019-03-29 | 2023-01-25 | Public University Corporation Yokohama City University | MATRIX COMPOSITION |
CN114341347A (zh) * | 2019-10-09 | 2022-04-12 | 国立大学法人大阪大学 | 血管内皮干细胞的制造方法 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013047639A1 (ja) | 2011-09-27 | 2013-04-04 | 公立大学法人横浜市立大学 | 組織及び臓器の作製方法 |
WO2015012158A1 (ja) * | 2013-07-23 | 2015-01-29 | 公立大学法人横浜市立大学 | 生物学的組織に血管系を付与する方法 |
JP5777127B1 (ja) | 2014-12-09 | 2015-09-09 | 公立大学法人横浜市立大学 | 原始腸内胚葉細胞及びその作製方法 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9677085B2 (en) * | 2013-03-18 | 2017-06-13 | Massachusetts Institute Of Technology | Engineering a heterogeneous tissue from pluripotent stem cells |
-
2016
- 2016-12-19 BR BR112018012070-7A patent/BR112018012070A2/ja not_active IP Right Cessation
- 2016-12-19 WO PCT/JP2016/087739 patent/WO2017110724A1/ja active Application Filing
- 2016-12-19 AU AU2016374909A patent/AU2016374909B2/en active Active
- 2016-12-19 CA CA3008213A patent/CA3008213A1/en not_active Abandoned
- 2016-12-19 EP EP16878613.5A patent/EP3395943B1/en active Active
- 2016-12-19 JP JP2017558110A patent/JP6860921B2/ja active Active
- 2016-12-19 SG SG11201804755VA patent/SG11201804755VA/en unknown
- 2016-12-19 KR KR1020187017472A patent/KR20180097564A/ko not_active Application Discontinuation
- 2016-12-19 CN CN201680070326.9A patent/CN108368484B/zh active Active
- 2016-12-19 US US16/060,814 patent/US20180352792A1/en not_active Abandoned
-
2018
- 2018-12-24 HK HK18116553.5A patent/HK1257345A1/zh unknown
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013047639A1 (ja) | 2011-09-27 | 2013-04-04 | 公立大学法人横浜市立大学 | 組織及び臓器の作製方法 |
WO2015012158A1 (ja) * | 2013-07-23 | 2015-01-29 | 公立大学法人横浜市立大学 | 生物学的組織に血管系を付与する方法 |
JP5777127B1 (ja) | 2014-12-09 | 2015-09-09 | 公立大学法人横浜市立大学 | 原始腸内胚葉細胞及びその作製方法 |
Non-Patent Citations (22)
Title |
---|
BILITY MT. ET AL., NAT PROTOC., vol. 7, 2012, pages 1608 - 17 |
D. ZHANG ET AL., CELL RES., vol. 19, no. 4, 2009, pages 429 - 38 |
G. LEE ET AL., NATURE BIOTECHNOLOGY, vol. 25, 2007, pages 1468 - 1475 |
HIDEKI TANIGUCHI ET AL.: "Generation of a vascularized human liver from an iPSC- derived organ bud transplant", KAN TAN SUI, vol. 70, no. 3, March 2015 (2015-03-01), pages 353 - 360, XP009511499 * |
HJ. JOO ET AL., BLOOD, vol. 118, no. 8, 2011, pages 2094 - 104 |
J. CAI ET AL., J MOL CELL BIOL., vol. 2, no. 1, 2010, pages 50 - 60 |
J. ZHANG ET AL., CIRCULATION RESEARCH., vol. 104, 2009, pages e30 - e41 |
JR. SPENCE ET AL., NATURE, vol. 470, no. 7332, 2011, pages 105 - 9 |
K. MATSUMOTO ET AL., SCIENCE, vol. 19, no. 5542, 2001, pages 559 - 63 |
K.SI-TAIYEB ET AL., HEPATOLOGY, vol. 51, no. 1, 2010, pages 297 - 305 |
KAMIYA A. ET AL., EMBO J., vol. 18, 1999, pages 2127 - 3 |
KINOSHITA T. ET AL., PROC NATL ACAD SCI USA., vol. 96, 1999, pages 7265 - 70 |
R. PETERS ET AL., PLOS ONE, vol. 5, no. 12, 2010, pages e15689 |
R. SPENCE ET AL., NATURE, vol. 470, no. 7332, 2011, pages 105 - 9 |
See also references of EP3395943A4 |
SELF ET AL., EMBO J., vol. 25, no. 21, 2006, pages 5214 - 5228 |
T. TOUBOUL ET AL., HEPATOLOGY, vol. 51, no. 5, 2010, pages 1754 - 65 |
TAKANORI TAKEBE ET AL.: "Creation of vascularized and functional organ from human iPS cell", ORGAN BIOLOGY, vol. 20, no. 2, 2013, pages 182 - 185, XP 009511496 * |
TAKEBE ET AL., NATURE, vol. 499, no. 7459, 2013, pages 481 - 484 |
TAKEBE T. ET AL., NATURE, 2013 |
TAKEBE, TAKANORI. ET AL.: "Generation of a vascularized and functional human liver from an iPSC-derived organ bud transplant", NATURE PROTOCOLS, vol. 9, no. 2, 23 January 2014 (2014-01-23), pages 396 - 409, XP 055166485 * |
WASHBURN ML. ET AL., GASTROENTEROLOGY, vol. 140, 2011, pages 1334 - 44 |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2019189324A1 (ja) * | 2018-03-28 | 2019-10-03 | 公立大学法人横浜市立大学 | 細胞塊融合法 |
CN111770989A (zh) * | 2018-03-28 | 2020-10-13 | 公立大学法人横滨市立大学 | 细胞团块融合方法 |
JPWO2019189324A1 (ja) * | 2018-03-28 | 2021-04-22 | 公立大学法人横浜市立大学 | 細胞塊融合法 |
JP7228269B2 (ja) | 2018-03-28 | 2023-02-24 | 公立大学法人横浜市立大学 | 細胞塊融合法 |
WO2020175594A1 (ja) * | 2019-02-28 | 2020-09-03 | 公立大学法人横浜市立大学 | 血液凝固および/または補体異常疾患の治療用組成物 |
CN114585746A (zh) * | 2019-10-17 | 2022-06-03 | 公立大学法人横滨市立大学 | 药物毒性评价方法 |
EP4047082A4 (en) * | 2019-10-17 | 2023-08-16 | Public University Corporation Yokohama City University | DRUG TOXICITY EVALUATION PROCEDURE |
Also Published As
Publication number | Publication date |
---|---|
SG11201804755VA (en) | 2018-07-30 |
BR112018012070A2 (ja) | 2018-11-27 |
EP3395943A4 (en) | 2019-08-28 |
KR20180097564A (ko) | 2018-08-31 |
JPWO2017110724A1 (ja) | 2018-10-11 |
AU2016374909A1 (en) | 2018-07-05 |
CA3008213A1 (en) | 2017-06-29 |
AU2016374909B2 (en) | 2022-10-06 |
CN108368484B (zh) | 2022-05-10 |
EP3395943B1 (en) | 2022-11-30 |
JP6860921B2 (ja) | 2021-04-21 |
CN108368484A (zh) | 2018-08-03 |
US20180352792A1 (en) | 2018-12-13 |
EP3395943A1 (en) | 2018-10-31 |
HK1257345A1 (zh) | 2019-10-18 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6860921B2 (ja) | 血液細胞を用いた組織・臓器の作製方法 | |
JP6455934B2 (ja) | 生物学的組織に血管系を付与する方法 | |
MXPA06006706A (es) | Celulas madre. | |
WO2012133948A1 (ja) | 生体組織から単離できるssea-3陽性の多能性幹細胞を含む他家移植用細胞治療用組成物 | |
JP2011519574A (ja) | 細胞ベースの治療に関連する材料および方法 | |
Melero‐Martin et al. | An in vivo experimental model for postnatal vasculogenesis | |
JP2014132830A (ja) | 生体の臍帯又は脂肪組織から単離できる多能性幹細胞 | |
JP7158040B2 (ja) | 構造体と細胞塊を連結した構築物 | |
Watanabe et al. | Bone marrow contributes to the population of pancreatic stellate cells in mice | |
US20100158874A1 (en) | Compositions and Methods for Treating Peripheral Vascular Diseases | |
CN108350418B (zh) | 病毒感染模型、其制作方法及其利用 | |
JP7228269B2 (ja) | 細胞塊融合法 | |
JP7385929B2 (ja) | 線維化を伴う疾患の予防及び/又は治療剤 | |
Chen et al. | Bioprinting of Human Cord Blood-Derived CD34+ Cells and Exploration of the Multilineage Differentiation Ability in Vitro | |
Petinati et al. | Multipotent Mesenchymal Stromal Cells from Porcine Bone Marrow, Implanted under the Kidney Capsule, form an Ectopic Focus Containing Bone, Hematopoietic Stromal Microenvironment, and Muscles. Cells 2023, 12, 268 | |
CA3179498A1 (en) | High-potential pluripotent stem cells | |
Watanabe et al. | Articles in PresS. Am J Physiol Gastrointest Liver Physiol (October 1, 2009). doi: 10.1152/ajpgi. 00123.2009 | |
JPWO2018003997A1 (ja) | 臓器線維症の予防または治療剤 | |
Chong | Defining the origin of adult cardiac multipotent colony forming stem cells | |
YOUNG | Isolation and characterization of human fetal liver progenitor stem cells | |
Howell | Identification of a common pluripotent stem cell population derived from multiple murine tissues |
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: 16878613 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 2017558110 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 11201804755V Country of ref document: SG |
|
ENP | Entry into the national phase |
Ref document number: 3008213 Country of ref document: CA |
|
ENP | Entry into the national phase |
Ref document number: 20187017472 Country of ref document: KR Kind code of ref document: A |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
REG | Reference to national code |
Ref country code: BR Ref legal event code: B01A Ref document number: 112018012070 Country of ref document: BR |
|
ENP | Entry into the national phase |
Ref document number: 2016374909 Country of ref document: AU Date of ref document: 20161219 Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2016878613 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 2016878613 Country of ref document: EP Effective date: 20180724 |
|
ENP | Entry into the national phase |
Ref document number: 112018012070 Country of ref document: BR Kind code of ref document: A2 Effective date: 20180614 |