JP7343984B2 - ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 - Google Patents
ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 Download PDFInfo
- Publication number
- JP7343984B2 JP7343984B2 JP2019022273A JP2019022273A JP7343984B2 JP 7343984 B2 JP7343984 B2 JP 7343984B2 JP 2019022273 A JP2019022273 A JP 2019022273A JP 2019022273 A JP2019022273 A JP 2019022273A JP 7343984 B2 JP7343984 B2 JP 7343984B2
- Authority
- JP
- Japan
- Prior art keywords
- cells
- hemangioblasts
- various embodiments
- item
- cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims description 96
- 210000003566 hemangioblast Anatomy 0.000 title claims description 82
- 210000000822 natural killer cell Anatomy 0.000 title claims description 55
- 210000001671 embryonic stem cell Anatomy 0.000 title claims description 20
- 210000004443 dendritic cell Anatomy 0.000 title description 93
- 210000004027 cell Anatomy 0.000 claims description 148
- 229920000609 methyl cellulose Polymers 0.000 claims description 34
- 239000001923 methylcellulose Substances 0.000 claims description 34
- 238000012258 culturing Methods 0.000 claims description 27
- 102000003812 Interleukin-15 Human genes 0.000 claims description 24
- 108090000172 Interleukin-15 Proteins 0.000 claims description 24
- 210000004263 induced pluripotent stem cell Anatomy 0.000 claims description 24
- 210000002966 serum Anatomy 0.000 claims description 21
- 210000001778 pluripotent stem cell Anatomy 0.000 claims description 15
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 claims description 9
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 claims description 9
- 239000001963 growth medium Substances 0.000 claims description 4
- 102100020715 Fms-related tyrosine kinase 3 ligand protein Human genes 0.000 claims 3
- 101710162577 Fms-related tyrosine kinase 3 ligand protein Proteins 0.000 claims 3
- 102000003951 Erythropoietin Human genes 0.000 claims 1
- 108090000394 Erythropoietin Proteins 0.000 claims 1
- 102000000704 Interleukin-7 Human genes 0.000 claims 1
- 108010002586 Interleukin-7 Proteins 0.000 claims 1
- 102100020880 Kit ligand Human genes 0.000 claims 1
- 101710177504 Kit ligand Proteins 0.000 claims 1
- 229940105423 erythropoietin Drugs 0.000 claims 1
- OXCMYAYHXIHQOA-UHFFFAOYSA-N potassium;[2-butyl-5-chloro-3-[[4-[2-(1,2,4-triaza-3-azanidacyclopenta-1,4-dien-5-yl)phenyl]phenyl]methyl]imidazol-4-yl]methanol Chemical compound [K+].CCCCC1=NC(Cl)=C(CO)N1CC1=CC=C(C=2C(=CC=CC=2)C2=N[N-]N=N2)C=C1 OXCMYAYHXIHQOA-UHFFFAOYSA-N 0.000 claims 1
- 102000004127 Cytokines Human genes 0.000 description 50
- 108090000695 Cytokines Proteins 0.000 description 50
- 239000000203 mixture Substances 0.000 description 39
- 239000002609 medium Substances 0.000 description 34
- 239000007788 liquid Substances 0.000 description 26
- 210000001082 somatic cell Anatomy 0.000 description 23
- 230000004069 differentiation Effects 0.000 description 22
- 230000008672 reprogramming Effects 0.000 description 19
- 238000003556 assay Methods 0.000 description 15
- 210000002242 embryoid body Anatomy 0.000 description 15
- 241000894007 species Species 0.000 description 15
- 239000000427 antigen Substances 0.000 description 12
- 102000036639 antigens Human genes 0.000 description 12
- 108091007433 antigens Proteins 0.000 description 12
- 230000035800 maturation Effects 0.000 description 11
- 102000006354 HLA-DR Antigens Human genes 0.000 description 10
- 108010058597 HLA-DR Antigens Proteins 0.000 description 10
- 210000001744 T-lymphocyte Anatomy 0.000 description 9
- 230000024245 cell differentiation Effects 0.000 description 9
- 238000004519 manufacturing process Methods 0.000 description 9
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 8
- 210000001185 bone marrow Anatomy 0.000 description 8
- 238000000684 flow cytometry Methods 0.000 description 8
- 230000001024 immunotherapeutic effect Effects 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 230000004044 response Effects 0.000 description 8
- 210000000130 stem cell Anatomy 0.000 description 8
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 7
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 7
- 238000003501 co-culture Methods 0.000 description 7
- XEYBRNLFEZDVAW-ARSRFYASSA-N dinoprostone Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1C\C=C/CCCC(O)=O XEYBRNLFEZDVAW-ARSRFYASSA-N 0.000 description 7
- 229960002986 dinoprostone Drugs 0.000 description 7
- 210000001939 mature NK cell Anatomy 0.000 description 7
- 230000004048 modification Effects 0.000 description 7
- 238000012986 modification Methods 0.000 description 7
- XEYBRNLFEZDVAW-UHFFFAOYSA-N prostaglandin E2 Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CC=CCCCC(O)=O XEYBRNLFEZDVAW-UHFFFAOYSA-N 0.000 description 7
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 6
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 6
- 102100022297 Integrin alpha-X Human genes 0.000 description 6
- 108091036414 Polyinosinic:polycytidylic acid Proteins 0.000 description 6
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 6
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 6
- 230000003013 cytotoxicity Effects 0.000 description 6
- 231100000135 cytotoxicity Toxicity 0.000 description 6
- 238000002825 functional assay Methods 0.000 description 6
- 230000003394 haemopoietic effect Effects 0.000 description 6
- 238000003306 harvesting Methods 0.000 description 6
- 210000003509 immature nk cell Anatomy 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 239000000546 pharmaceutical excipient Substances 0.000 description 6
- 229940115272 polyinosinic:polycytidylic acid Drugs 0.000 description 6
- 102100021992 CD209 antigen Human genes 0.000 description 5
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 5
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 description 5
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 description 5
- 102000004142 Trypsin Human genes 0.000 description 5
- 108090000631 Trypsin Proteins 0.000 description 5
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 5
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 5
- 208000021841 acute erythroid leukemia Diseases 0.000 description 5
- 230000006907 apoptotic process Effects 0.000 description 5
- 238000002784 cytotoxicity assay Methods 0.000 description 5
- 231100000263 cytotoxicity test Toxicity 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 5
- 239000012588 trypsin Substances 0.000 description 5
- 102100035793 CD83 antigen Human genes 0.000 description 4
- 241000282472 Canis lupus familiaris Species 0.000 description 4
- 241000283153 Cetacea Species 0.000 description 4
- 241000283086 Equidae Species 0.000 description 4
- 241000282326 Felis catus Species 0.000 description 4
- 101000946856 Homo sapiens CD83 antigen Proteins 0.000 description 4
- 101001109501 Homo sapiens NKG2-D type II integral membrane protein Proteins 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 102100022680 NKG2-D type II integral membrane protein Human genes 0.000 description 4
- 238000010817 Wright-Giemsa staining Methods 0.000 description 4
- 239000002458 cell surface marker Substances 0.000 description 4
- 210000004748 cultured cell Anatomy 0.000 description 4
- 230000001472 cytotoxic effect Effects 0.000 description 4
- 230000001419 dependent effect Effects 0.000 description 4
- 210000002950 fibroblast Anatomy 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 229940092253 ovalbumin Drugs 0.000 description 4
- 239000008194 pharmaceutical composition Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000028327 secretion Effects 0.000 description 4
- 238000002560 therapeutic procedure Methods 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 239000013598 vector Substances 0.000 description 4
- 102100024505 Bone morphogenetic protein 4 Human genes 0.000 description 3
- -1 CD16 Proteins 0.000 description 3
- 208000031637 Erythroblastic Acute Leukemia Diseases 0.000 description 3
- 208000036566 Erythroleukaemia Diseases 0.000 description 3
- 241000282412 Homo Species 0.000 description 3
- 101000762379 Homo sapiens Bone morphogenetic protein 4 Proteins 0.000 description 3
- 101000971513 Homo sapiens Natural killer cells antigen CD94 Proteins 0.000 description 3
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 3
- 102000003810 Interleukin-18 Human genes 0.000 description 3
- 108090000171 Interleukin-18 Proteins 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- 102100021462 Natural killer cells antigen CD94 Human genes 0.000 description 3
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 210000002459 blastocyst Anatomy 0.000 description 3
- 231100000433 cytotoxic Toxicity 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 210000002889 endothelial cell Anatomy 0.000 description 3
- 230000001605 fetal effect Effects 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 238000009630 liquid culture Methods 0.000 description 3
- 210000001161 mammalian embryo Anatomy 0.000 description 3
- 238000007799 mixed lymphocyte reaction assay Methods 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 230000008186 parthenogenesis Effects 0.000 description 3
- PHEDXBVPIONUQT-RGYGYFBISA-N phorbol 13-acetate 12-myristate Chemical compound C([C@]1(O)C(=O)C(C)=C[C@H]1[C@@]1(O)[C@H](C)[C@H]2OC(=O)CCCCCCCCCCCCC)C(CO)=C[C@H]1[C@H]1[C@]2(OC(C)=O)C1(C)C PHEDXBVPIONUQT-RGYGYFBISA-N 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 210000002536 stromal cell Anatomy 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- 241001455214 Acinonyx jubatus Species 0.000 description 2
- 241000282452 Ailuropoda melanoleuca Species 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 108010077544 Chromatin Proteins 0.000 description 2
- 241001125840 Coryphaenidae Species 0.000 description 2
- 241000699800 Cricetinae Species 0.000 description 2
- 102000001398 Granzyme Human genes 0.000 description 2
- 108060005986 Granzyme Proteins 0.000 description 2
- 101000973177 Homo sapiens Nuclear factor interleukin-3-regulated protein Proteins 0.000 description 2
- 101000914496 Homo sapiens T-cell antigen CD7 Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 102000014736 Notch Human genes 0.000 description 2
- 108010070047 Notch Receptors Proteins 0.000 description 2
- 102100022163 Nuclear factor interleukin-3-regulated protein Human genes 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 208000025174 PANDAS Diseases 0.000 description 2
- 208000021155 Paediatric autoimmune neuropsychiatric disorders associated with streptococcal infection Diseases 0.000 description 2
- 240000004718 Panda Species 0.000 description 2
- 235000016496 Panda oleosa Nutrition 0.000 description 2
- 241000282320 Panthera leo Species 0.000 description 2
- 241000282376 Panthera tigris Species 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- KHGNFPUMBJSZSM-UHFFFAOYSA-N Perforine Natural products COC1=C2CCC(O)C(CCC(C)(C)O)(OC)C2=NC2=C1C=CO2 KHGNFPUMBJSZSM-UHFFFAOYSA-N 0.000 description 2
- 241000283080 Proboscidea <mammal> Species 0.000 description 2
- 241000282335 Procyon Species 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 101100247004 Rattus norvegicus Qsox1 gene Proteins 0.000 description 2
- 241000283984 Rodentia Species 0.000 description 2
- 241000282887 Suidae Species 0.000 description 2
- 102100027208 T-cell antigen CD7 Human genes 0.000 description 2
- 102100036011 T-cell surface glycoprotein CD4 Human genes 0.000 description 2
- 206010043276 Teratoma Diseases 0.000 description 2
- 210000005006 adaptive immune system Anatomy 0.000 description 2
- 239000000443 aerosol Substances 0.000 description 2
- 230000000735 allogeneic effect Effects 0.000 description 2
- 238000011319 anticancer therapy Methods 0.000 description 2
- 210000003969 blast cell Anatomy 0.000 description 2
- 210000001109 blastomere Anatomy 0.000 description 2
- 239000002771 cell marker Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 210000003483 chromatin Anatomy 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 239000012636 effector Substances 0.000 description 2
- 210000002257 embryonic structure Anatomy 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000004720 fertilization Effects 0.000 description 2
- 210000004700 fetal blood Anatomy 0.000 description 2
- 210000001654 germ layer Anatomy 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 210000005007 innate immune system Anatomy 0.000 description 2
- 230000000366 juvenile effect Effects 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 210000003071 memory t lymphocyte Anatomy 0.000 description 2
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 210000000472 morula Anatomy 0.000 description 2
- 229930192851 perforin Natural products 0.000 description 2
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 230000002285 radioactive effect Effects 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 230000001177 retroviral effect Effects 0.000 description 2
- 230000011664 signaling Effects 0.000 description 2
- 210000000329 smooth muscle myocyte Anatomy 0.000 description 2
- 238000010374 somatic cell nuclear transfer Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 230000032258 transport Effects 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 229960005486 vaccine Drugs 0.000 description 2
- 230000002792 vascular Effects 0.000 description 2
- 210000005167 vascular cell Anatomy 0.000 description 2
- 208000030507 AIDS Diseases 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 1
- 208000011691 Burkitt lymphomas Diseases 0.000 description 1
- 102100036842 C-C motif chemokine 19 Human genes 0.000 description 1
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 1
- 108050005493 CD3 protein, epsilon/gamma/delta subunit Proteins 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- 108010037897 DC-specific ICAM-3 grabbing nonintegrin Proteins 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000699694 Gerbillinae Species 0.000 description 1
- 102100035716 Glycophorin-A Human genes 0.000 description 1
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 1
- 101000713106 Homo sapiens C-C motif chemokine 19 Proteins 0.000 description 1
- 101001074244 Homo sapiens Glycophorin-A Proteins 0.000 description 1
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 1
- 101000716102 Homo sapiens T-cell surface glycoprotein CD4 Proteins 0.000 description 1
- 101000687905 Homo sapiens Transcription factor SOX-2 Proteins 0.000 description 1
- 101000642523 Homo sapiens Transcription factor SOX-7 Proteins 0.000 description 1
- 101000976622 Homo sapiens Zinc finger protein 42 homolog Proteins 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010061598 Immunodeficiency Diseases 0.000 description 1
- 102000013462 Interleukin-12 Human genes 0.000 description 1
- 108010065805 Interleukin-12 Proteins 0.000 description 1
- 102000002698 KIR Receptors Human genes 0.000 description 1
- 108010043610 KIR Receptors Proteins 0.000 description 1
- 108700021430 Kruppel-Like Factor 4 Proteins 0.000 description 1
- 108010013709 Leukocyte Common Antigens Proteins 0.000 description 1
- 102000017095 Leukocyte Common Antigens Human genes 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- 239000005089 Luciferase Substances 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 102000043129 MHC class I family Human genes 0.000 description 1
- 108091054437 MHC class I family Proteins 0.000 description 1
- 102000043131 MHC class II family Human genes 0.000 description 1
- 108091054438 MHC class II family Proteins 0.000 description 1
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 1
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 1
- 102100038895 Myc proto-oncogene protein Human genes 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
- 102000001708 Protein Isoforms Human genes 0.000 description 1
- 108010029485 Protein Isoforms Proteins 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 238000011579 SCID mouse model Methods 0.000 description 1
- 101150086694 SLC22A3 gene Proteins 0.000 description 1
- 241000555745 Sciuridae Species 0.000 description 1
- 108091008035 T cell costimulatory receptors Proteins 0.000 description 1
- 102100024270 Transcription factor SOX-2 Human genes 0.000 description 1
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 1
- 102100033177 Vascular endothelial growth factor receptor 2 Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 102100023550 Zinc finger protein 42 homolog Human genes 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 208000030961 allergic reaction Diseases 0.000 description 1
- 210000000648 angioblast Anatomy 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 230000029918 bioluminescence Effects 0.000 description 1
- 238000005415 bioluminescence Methods 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000002798 bone marrow cell Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 210000004413 cardiac myocyte Anatomy 0.000 description 1
- 230000011712 cell development Effects 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000002975 chemoattractant Substances 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 230000016396 cytokine production Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 210000002308 embryonic cell Anatomy 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 230000000925 erythroid effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 1
- 210000003953 foreskin Anatomy 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 210000002064 heart cell Anatomy 0.000 description 1
- 102000052241 human SOX7 Human genes 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 238000013394 immunophenotyping Methods 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229940102223 injectable solution Drugs 0.000 description 1
- 229940102213 injectable suspension Drugs 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000007913 intrathecal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- PGHMRUGBZOYCAA-ADZNBVRBSA-N ionomycin Chemical compound O1[C@H](C[C@H](O)[C@H](C)[C@H](O)[C@H](C)/C=C/C[C@@H](C)C[C@@H](C)C(/O)=C/C(=O)[C@@H](C)C[C@@H](C)C[C@@H](CCC(O)=O)C)CC[C@@]1(C)[C@@H]1O[C@](C)([C@@H](C)O)CC1 PGHMRUGBZOYCAA-ADZNBVRBSA-N 0.000 description 1
- PGHMRUGBZOYCAA-UHFFFAOYSA-N ionomycin Natural products O1C(CC(O)C(C)C(O)C(C)C=CCC(C)CC(C)C(O)=CC(=O)C(C)CC(C)CC(CCC(O)=O)C)CCC1(C)C1OC(C)(C(C)O)CC1 PGHMRUGBZOYCAA-UHFFFAOYSA-N 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 101150111214 lin-28 gene Proteins 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 238000010232 migration assay Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 210000000581 natural killer T-cell Anatomy 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000011176 pooling Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000002629 repopulating effect Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 210000005222 synovial tissue Anatomy 0.000 description 1
- 210000002437 synoviocyte Anatomy 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 238000012447 xenograft mouse model Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0639—Dendritic cells, e.g. Langherhans cells in the epidermis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/15—Cells of the myeloid line, e.g. granulocytes, basophils, eosinophils, neutrophils, leucocytes, monocytes, macrophages or mast cells; Myeloid precursor cells; Antigen-presenting cells, e.g. dendritic cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/461—Cellular immunotherapy characterised by the cell type used
- A61K39/4613—Natural-killer cells [NK or NK-T]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/46—Cellular immunotherapy
- A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
- A61K39/4643—Vertebrate antigens
- A61K39/4644—Cancer antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- 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
- C12N5/0646—Natural killers cells [NK], NKT cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K2035/124—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells the cells being hematopoietic, bone marrow derived or blood cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K39/46
- A61K2239/46—Indexing codes associated with cellular immunotherapy of group A61K39/46 characterised by the cancer treated
- A61K2239/48—Blood cells, e.g. leukemia or lymphoma
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/125—Stem cell factor [SCF], c-kit ligand [KL]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/22—Colony stimulating factors (G-CSF, GM-CSF)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2303—Interleukin-3 (IL-3)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2304—Interleukin-4 (IL-4)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/26—Flt-3 ligand (CD135L, flk-2 ligand)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
- C12N2506/02—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from embryonic cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oncology (AREA)
- Developmental Biology & Embryology (AREA)
- Virology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Description
例えば、本発明は以下を提供する。
(項目1)
ナチュラルキラー(NK)細胞を生成する方法であって、
血管芽細胞を提供すること、
前記血管芽細胞を、メチルセルロース、並びにIL2、IL3、IL6、IL7、IL15、SCF、およびFLを含む第1のサイトカイン混合物上で培養すること、
前記培養した細胞を回収すること、および
前記回収した細胞を、ヒト血清、並びにIL7、IL15、SCF、およびFLを含む第2のサイトカイン混合物を含む液体培地中で培養してNK細胞を生成することを含む方法。
(項目2)
前記メチルセルロースが、H4236メチルセルロースである、項目1に記載の方法。(項目3)
前記メチルセルロースが、H4536メチルセルロースである、項目1に記載の方法。(項目4)
IL2の濃度が、約5~10ng/mlであり、IL3が約1~10ng/mlであり、IL6が約1~10ng/mlであり、IL7が約5~20ng/mlであり、IL15が約5~10ng/mlであり、SCFが約10~50ng/mlであり、FLが約10~50ng/mlである、項目1に記載の方法。
(項目5)
前記血管芽細胞の培養が、約6~8日間である、項目1に記載の方法。
(項目6)
前記回収した細胞の培養が、約14~21日間である、項目1に記載の方法。
(項目7)
培地を週1回変更して、前記第2のサイトカイン混合物を新しくすることを更に含む、項目1に記載の方法。
(項目8)
前記血管芽細胞が、ヒト胚性幹細胞(hESC)から分化する、項目1に記載の方法。(項目9)
前記血管芽細胞が、誘導多能性(iPS)細胞から分化する、項目1に記載の方法。
(項目10)
前記NK細胞が、未熟NK細胞であり、CD56+およびCD16-である、項目1に記載の方法。
(項目11)
前記NK細胞が、成熟NK細胞であり、CD56-およびCD16+であるか、またはCD56loおよびCD16+である、項目1に記載の方法。
(項目12)
項目1~11のいずれか一項に記載の方法により生成されるナチュラルキラー(NK)細胞。
(項目13)
項目12に記載のNK細胞をある量含む薬学的に許容される組成物。
(項目14)
ナチュラルキラー(NK)細胞を生成する方法であって、
血管芽細胞を提供すること、
前記血管芽細胞を、ヒト血清、並びにIL2、IL3、IL6、IL7、IL15、およびSCFを含む第1のサイトカイン混合物を含む液体培地中で培養すること、
前記培養した細胞を回収すること、および
前記回収した細胞を、ヒト血清、並びにIL7、IL15、SCF、およびFLを含む第2のサイトカイン混合物を含む液体培地中で培養して前記NKを生成することを含む方法。
(項目15)
IL2の濃度が、約5~10ng/mlであり、IL3が約1~10ng/mlであり、IL6が約1~10ng/mlであり、IL7が約5~20ng/mlであり、IL15が約5~10ng/mlであり、SCFが約10~50ng/mlであり、FLが約10~50ng/mlである、項目14に記載の方法。
(項目16)
前記血管芽細胞の培養が、約6~8日間ある、項目14に記載の方法。
(項目17)
前記回収した細胞の培養が、約14~21日間である、項目14に記載の方法。
(項目18)
培地を週1回変更して、前記第2のサイトカイン混合物を新しくすることを更に含む、項目14に記載の方法。
(項目19)
前記血管芽細胞が、ヒト胚性幹細胞(hESC)から分化する、項目14に記載の方法。
(項目20)
前記血管芽細胞が、誘導多能性(iPS)細胞から分化する、項目14に記載の方法。(項目21)
前記NK細胞が、未熟NK細胞であり、CD56+およびCD16-である、項目14に記載の方法。
(項目22)
前記NK細胞が、成熟NK細胞であり、CD56-およびCD16+であるか、またはCD56loおよびCD16+である、項目14に記載の方法。
(項目23)
項目14~22のいずれか一項に記載の方法により生成されるナチュラルキラー(NK)細胞。
(項目24)
項目24に記載のNK細胞をある量含む薬学的に許容される組成物。
(項目25)
樹状細胞(DC)を生成する方法であって、
血管芽細胞を提供すること、
前記血管芽細胞を、ヒト血清、SCF、FL、IL3、およびGM-CSFを含む液体培地中で培養すること、
前記液体培地にIL4を添加すること、および
前記血管芽細胞を更に培養して、前記DCを生成することを含む方法。
(項目26)
前記血管芽細胞の培養が、約7~11日間ある、項目25に記載の方法。
(項目27)
前記IL4添加の後の前記血管芽細胞の培養が、約8~10日間である、項目25に記載の方法。
(項目28)
IL1b、TNFα、およびIL6を含むサイトカイン混合物を添加して、前記DCの成熟化を誘導することを更に含む、項目25に記載の方法。
(項目29)
前記サイトカイン混合物が、約48時間添加される、項目28に記載の方法。
(項目30)
前記サイトカイン混合物が、PGE2、IFNα2b、Poly I:C、IFNγ、およびそれらの組み合わせからなる群から選択されるサイトカインを更に含む、項目28に記載の方法。
(項目31)
LPS、IFNγ、および/またはS-28463を添加して、前記DCからのIL12p70産生および/または前記DCからのHLA-DR発現を刺激することを更に含む、項目25に記載の方法。
(項目32)
SCFの濃度が、約20~100ng/mlであり、FLが約10~50ng/mlであり、IL3が約5~50ng/mlであり、GM-CSFが約50~100ng/mlであり、IL4が約50~100ng/mlである、項目25に記載の方法。
(項目33)
IL1bの濃度が、約10ng/mlであり、TNFγが、約10ng/mlであり、IL6が約150ng/mlである、項目28に記載の方法。
(項目34)
PGE2の濃度が、約1μg/mlであり、IFNα2bが約3000単位/mlであり、poly I:Cが約20μg/mlであり、IFNγが約20ng/mlである、項目30に記載の方法。
(項目35)
前記DCが、成熟DCであり、CD83を発現する、項目25に記載の方法。
(項目36)
前記DCが、成熟DCであり、CD209、HLA DR、および/またはCD11cの発現が増加される、項目25に記載の方法。
(項目37)
項目25~36のいずれか一項に記載の方法により生成される樹状細胞(DC)細胞。(項目38)
項目37に記載のDC細胞をある量含む薬学的に許容される組成物。
初期分化
両細胞タイプの初期分化手順は同じであり、胚様体(EB)を生成するために、サイトカインを加えたStemline II(Sigma社製)中でhESCを4日間培養することを伴う。サイトカイン、VEGFおよびBMP4を、EB培養の全体にわたって使用し、bFGFを、最初の2日間の後に添加する。合計で4日後に、その結果生じたEBを、0.05%トリプシンで脱凝集し、その後トリプシンを血清含有培地で不活化する。その後、個々の細胞を40μM細胞ストレーナーでろ過し、計数し、TPO(50μg/ml)、VEGF(50μg/ml)、FL(50μg/ml)、およびbFGF(20~50μg/ml)等のさらなるサイトカインを含有するH4436またはH4536メチルセルロース(Stem Cell Technologies社製)に播種する。NK分化の場合、サイトカインIL2(1~10μg/ml)、IL7(1~20μg/ml)、および/またはIL15(1~10μg/ml)を、この段階で添加することもできる。血管芽細胞集団を産生および増殖させるために、1ml当たり50,000~150,000細胞の濃度で、メチルセルロース培養物を播種する。芽球様細胞を、6~10日目にメチルセルロースから回収し、下記の手順のうちの1つにより更に分化させる。
NK分化
芽球細胞は、IL2(5~10ng/ml)、IL3(1~10ng/ml)、IL6(1~10ng/ml)、IL7(5~20ng/ml)、IL15(5~10ng/ml)、SCF(10~50ng/ml)、およびFL(10~50ng/ml)を加えたH4236メチルセルロースに再播種してもよく、または同じサイトカインおよび10~20%ヒト血清を含有する液体培養に再播種してもよい。6~8日間の培養後、細胞を回収し、10~20%ヒト血清並びにサイトカインIL7(5~20ng/ml)、IL15(5~10ng/ml)、SCF(10~50ng/ml)、およびFL(10~50ng/ml)を加えた液体培地(αMEMまたはDMEM:F12)に、更に14~21日間再播種する。週1回の培地変更を使用して、サイトカイン混合物を新しくする。
DC分化
芽球細胞を、10~20%ヒト血清並びにサイトカイン、SCF(20~100ng/ml)、FL(10~50ng/ml)、IL3(5~50ng/ml)、およびGM-CSF(50~100ng/ml)を加えた液体培地(αMEMまたはDMEM:F12)に播種する。7~11日間の培養後、IL4(50~100ng/ml)も培養物に添加し、細胞を更に8~10日間増殖させる。培地変更は、6~7日毎に実施する。DCの成熟化を誘導するために、さらなるサイトカイン混合物(10ng/ml IL1b、10ng/ml TNFγ、150ng/ml IL6)を、48時間培養物に添加することができる。
血管芽細胞増殖条件の変更
H4436メチルセルロースで増殖させた血管芽細胞を使用して、上記のNKおよびDC分化手順を実施した。しかしながら、H4536、Stem Cell Technologies社製の無エリトロポイエチンメチルセルロースを使用して、血管芽細胞を効率的に生成することもできる。これら「epoマイナス」血管芽細胞は、元々の「epoプラス」芽球と非常に類似しており、それらは、様々な造血細胞タイプおよび血管細胞タイプへと分化することが可能である。予備的な結果は、NK細胞およびDCを含む種々の造血系列へと血管芽細胞を分化させるためには、H4536の使用が、H4436メチルセルロースよりも著しい利点を提供することができることを示唆する。芽球増殖培地中にepoが存在しないことにより、赤血球マーカーCD235aを発現する細胞の割合を低減させ、CD34、CD45、およびCD41aを発現する細胞の割合を増加させることが見出された。細胞表面マーカー発現におけるこの違いのため、「epo-マイナス」増殖条件は、骨髄系列および/またはリンパ系列への分化を増強することができる。
iPS細胞からのMKの生成
OP9共培養系を使用して、本発明者らは、iPS細胞からMKを生成することができることを示す。数千個のCD41a+MKを、数十万個のiPS細胞から生成した。
ヒトESCからのNK細胞分化
分化手順を以下のように実施した:
H7 ESCを、胚様体(EB)へと4日間分化させた。EBを回収し、血管芽細胞を産生および増殖させるために、サイトカイン豊富なメチルセルロースに10~15日間移した。血管芽細胞を回収し、10~20%ヒトAB血清を加えた無フィーダー液体培地に一群のサイトカインと共に更に14~17日間配置し、3~4日毎に培地を半分変更した。
免疫表現型検査(フローサイトメトリーを使用):
未熟NK細胞は、CD56bright、CD16lo、KIRlo、CD117+、CD94-、NKG2D+だった。上記の手順の変法を使用することにより、32日間の分化後の生細胞の20~30%が、CD56+CD16-だった。成熟NK細胞は、CD56dim、CD16hi、KIRhi、CD117lo/-、CD94+、NKG2D+だった。上記の手順を使用することにより、31日間の分化後の生細胞の20%が、CD56-CD16+であり、それらの5%が、CD56loCD16+だった。
ナチュラル細胞傷害性:成熟NK細胞は、ヒトK562赤白血病細胞、MCF7細胞、U87細胞、PC3細胞、NTERA2細胞等の標的細胞のアポトーシスを誘発することができる。「3FC」アッセイを使用して、細胞傷害性の効率を評価する。このアッセイは、51Cr放出アッセイと類似しているが、放射能を必要としない。Derby et al., Immunol. Letters 78: 35-39 (2001)を参照されたい。上述の成熟NK細胞の異種性集団(アイテムB2-a)が、標準的な4時間の実験で65~70%のK562細胞にアポトーシスを誘発することが見出された。
血管芽細胞を骨髄再増殖細胞として使用することにより、またはそれらを樹状突起細胞、ナチュラルキラー細胞、T細胞、および/または間葉系幹細胞(MSC)へと分化させることにより、本発明者らは、癌、HIV、および/または自己免疫疾患(automimmune disease)と闘うための大規模で効果的な細胞に基づく療法を生み出すことができる。本発明者らは、hESCおよびiPS細胞の両方からの樹状細胞(DC)の分化を、40~55%の効率で達成することができた。2つの新しい機能アッセイに加えて、ヒト骨髄に由来するDCと対照比較することにより、今や、hESC由来DCが、多くの同様な特徴をヒトBM由来DCと共有していることが確認され、さらなる最適化を必要とする分野も特定された。ナチュラルキラー細胞分化の場合、ヒト骨髄由来NK細胞と対照比較することにより、骨髄細胞供給源を使用した場合さえ、生体外NK細胞分化が、あまり効率的ではないことが確認された。本発明者らは、芽球由来NK細胞が、ナチュラル細胞傷害性能力を示すことを見出した。抗体依存性細胞傷害アッセイを実施する。T細胞分化の場合、本発明者らは、Notchシグナル伝達を刺激するためにヒトデルタリガンド発現OP9間質細胞系統の生成することに成功し、血管芽細胞のT細胞分化を刺激するために、この間質細胞系統を使用している。
細胞表面マーカー:CD11c、45、209は、芽球由来DCおよびヒト骨髄由来DCでは同様の発現を示すが、HLA-DRは、芽球DCでは、ヒトBM DCよりもはるかに低いレベルで発現される。
臍帯血および末梢血単核細胞は両方とも、レスポンダーとして使用される。本発明者らは、陽性対照エフェクターとしてヒト骨髄由来DCを使用する。
1. Woll, P, et al. J. of Immunology. 175: 5095-5103 (2005).
2. Woll, P et al. Blood113(24): 6094-101 (2009).
3. Bordoni et al. Hepatology 39: 1508-1516 (2004).
4. Tabatoabaei-Zavareh et al. PLOS One Issue 2, e232 (2007).
5. McCullar, V et al. Exp. Hematology 36(5): 598-608 (2008).
6. Freud, AG et al. Immunity 22:295-304 (2005).
7. Yu, H et al. Blood 92 (10): 3647-3657 (1998).
1. Su et al. Clinical Cancer Research 14(19): 6207-6217 (2008).
2. Tseng et al. Regenerative Medicine 4(4): 513-526 (2009).
3. Bandi et al. AIDS Research and Therapy 5:1 (2008). (open-access)
4. Slukvin, II et al. J of Immunology 176: 2924-2932 (2006).
Claims (7)
- ナチュラルキラー(NK)細胞を生成する方法であって、
無フィーダー条件で、メチルセルロース、SCF、Flt3リガンド(FL)、IL2、IL6、IL7、IL15およびIL3を含む第1の培養培地中で多能性幹細胞由来血管芽細胞を培養する工程、前記培養した細胞を回収する工程、および、前記回収した細胞を、無フィーダー条件で、ヒト血清、IL7、IL15、SCFおよびFLを含む第2の培養培地中で培養して、NK細胞を生成する工程を含む、方法。 - 前記多能性幹細胞が、ヒト胚性幹細胞またはヒト誘導多能性幹細胞である、請求項1に記載の方法。
- 前記血管芽細胞が、無フィーダー条件での多能性幹細胞から得られる、請求項1に記載の方法。
- 前記血管芽細胞が、エリトロポイエチンを用いる培養なしに、多能性幹細胞から得られる、請求項1に記載の方法。
- 前記NK細胞が、CD56+である、請求項1に記載の方法。
- 前記NK細胞が、CD56-である、請求項1に記載の方法。
- 前記第1の培養培地が、無血清である、請求項1に記載の方法。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2021188673A JP2022027790A (ja) | 2009-12-04 | 2021-11-19 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US26666109P | 2009-12-04 | 2009-12-04 | |
US61/266,661 | 2009-12-04 | ||
JP2016060101A JP2016119910A (ja) | 2009-12-04 | 2016-03-24 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2016060101A Division JP2016119910A (ja) | 2009-12-04 | 2016-03-24 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2021188673A Division JP2022027790A (ja) | 2009-12-04 | 2021-11-19 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2019083819A JP2019083819A (ja) | 2019-06-06 |
JP7343984B2 true JP7343984B2 (ja) | 2023-09-13 |
Family
ID=44115272
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2012542163A Active JP6013187B2 (ja) | 2009-12-04 | 2010-12-01 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
JP2016060101A Pending JP2016119910A (ja) | 2009-12-04 | 2016-03-24 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
JP2019022273A Active JP7343984B2 (ja) | 2009-12-04 | 2019-02-12 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
JP2021188673A Pending JP2022027790A (ja) | 2009-12-04 | 2021-11-19 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
Family Applications Before (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2012542163A Active JP6013187B2 (ja) | 2009-12-04 | 2010-12-01 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
JP2016060101A Pending JP2016119910A (ja) | 2009-12-04 | 2016-03-24 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2021188673A Pending JP2022027790A (ja) | 2009-12-04 | 2021-11-19 | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 |
Country Status (11)
Country | Link |
---|---|
US (3) | US8822218B2 (ja) |
EP (1) | EP2507365A4 (ja) |
JP (4) | JP6013187B2 (ja) |
KR (4) | KR20190016602A (ja) |
CN (1) | CN102822332A (ja) |
AU (1) | AU2010326027A1 (ja) |
BR (1) | BR112012014104A2 (ja) |
CA (3) | CA2781969A1 (ja) |
IN (1) | IN2012DN05053A (ja) |
RU (1) | RU2012127811A (ja) |
WO (1) | WO2011068896A1 (ja) |
Families Citing this family (28)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102651433B1 (ko) | 2006-04-14 | 2024-03-25 | 아스텔라스 인스티튜트 포 리제너러티브 메디슨 | 혈관 콜로니 형성 세포 |
BR112012014104A2 (pt) | 2009-12-04 | 2016-07-05 | Stem Cell & Regenerative Medicine International Inc | método de geração de células destrutoras naturais e células dendríticas de hemangioblastos embrionários humanos derivados de células-tronco |
EP3567102A1 (en) * | 2012-04-24 | 2019-11-13 | Dan S. Kaufman | Method for developing natural killer cells from stem cells |
CA2896053A1 (en) | 2012-12-21 | 2014-06-26 | Ocata Therapeutics, Inc. | Methods for production of platelets from pluripotent stem cells and compositions thereof |
CA2935459C (en) * | 2014-01-03 | 2021-04-27 | FullHope Biomedical Co., Ltd. | Modified natural killer cells, compositions and uses thereof |
CN105219710B (zh) * | 2014-06-05 | 2020-01-10 | 上海厚超生物科技有限公司 | 一种高杀伤活性的免疫细胞群的培养方法 |
DE102014014993B4 (de) * | 2014-10-09 | 2017-02-02 | Helga Schmetzer | Verwendung von immunmodulatorisch wirksamen Kits zur immuntherapeutischen Behandlung von Patienten mit myeloischen Leukämien |
CN104711225B (zh) * | 2015-04-09 | 2018-05-22 | 奥思达干细胞有限公司 | Nk细胞的体外制备方法 |
TWI772270B (zh) | 2015-08-18 | 2022-08-01 | 美商安斯泰來再生醫藥協會 | 臨床調配物 |
CN105219717A (zh) * | 2015-10-20 | 2016-01-06 | 上海隆耀生物科技有限公司 | 一种一型极化树突状细胞及其诱导方法和应用 |
CN105219726A (zh) * | 2015-10-20 | 2016-01-06 | 上海隆耀生物科技有限公司 | 一种高效制备一型极化树突状细胞的方法及其应用 |
CN105219728A (zh) * | 2015-10-20 | 2016-01-06 | 上海隆耀生物科技有限公司 | 一种用于激活乳腺癌特异性免疫反应的试剂盒 |
CN105219723A (zh) * | 2015-10-20 | 2016-01-06 | 上海隆耀生物科技有限公司 | 一种用于激活胃癌特异性免疫反应的试剂盒 |
CN105219716A (zh) * | 2015-10-20 | 2016-01-06 | 上海隆耀生物科技有限公司 | 一种用于激活头颈部癌特异性免疫反应的试剂盒 |
CN105219727A (zh) * | 2015-10-20 | 2016-01-06 | 上海隆耀生物科技有限公司 | 一种用于激活结直肠癌特异性免疫反应的试剂盒 |
CN105219722A (zh) * | 2015-10-20 | 2016-01-06 | 上海隆耀生物科技有限公司 | 一种用于激活肾癌特异性免疫反应的试剂盒 |
CN105219718A (zh) * | 2015-10-20 | 2016-01-06 | 上海隆耀生物科技有限公司 | 一种用于激活前列腺癌特异性免疫反应的试剂盒 |
CN105219725A (zh) * | 2015-10-20 | 2016-01-06 | 上海隆耀生物科技有限公司 | 一种用于激活肉瘤特异性免疫反应的试剂盒 |
CN105219719A (zh) * | 2015-10-20 | 2016-01-06 | 上海隆耀生物科技有限公司 | 一种用于激活卵巢癌特异性免疫反应的试剂盒 |
GB201810486D0 (en) * | 2018-06-26 | 2018-08-08 | Imperial Innovations Ltd | Natural killer cells |
US11453862B2 (en) | 2019-07-08 | 2022-09-27 | Immunitybio, Inc. | Mononuclear cell derived NK cells |
WO2021006875A1 (en) | 2019-07-08 | 2021-01-14 | Nantkwest, Inc. | Mononuclear cell derived nk cells |
US11547726B2 (en) * | 2020-03-26 | 2023-01-10 | Honed Life Sciences, Llc | Enhancement of production of NK cells from stem cells |
EP4251741A1 (en) | 2020-11-30 | 2023-10-04 | CRISPR Therapeutics AG | Gene-edited natural killer cells |
KR102292843B1 (ko) * | 2020-12-29 | 2021-08-25 | 주식회사 온코인사이트 | 역분화줄기세포(iPSC) 유래 자연 살해 세포 및 이의 용도 |
WO2022144632A1 (en) | 2020-12-30 | 2022-07-07 | Crispr Therapeutics Ag | Compositions and methods for differentiating stem cells into nk cells |
CN115247149B (zh) * | 2022-08-22 | 2023-06-16 | 华域生物科技(天津)有限公司 | 适用于nk细胞的培养基组合物及培养方法 |
WO2024047368A1 (en) * | 2022-09-02 | 2024-03-07 | Imperial College Innovations Limited | Natural killer cells |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070141703A1 (en) | 2003-11-19 | 2007-06-21 | Stanley Edouard G | Methods for producing blood products from pluripotent cells in cell culture |
WO2008056779A1 (fr) | 2006-11-09 | 2008-05-15 | Japan As Represented By The President Of International Medical Center Of Japan | Procédé destiné à la culture et au passage d'une cellule souche embryonnaire de primate, et procédé destiné à induire la différenciation de la cellule souche embryonnaire |
Family Cites Families (26)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ES2201102T3 (es) * | 1994-06-15 | 2004-03-16 | Systemix, Inc. | Poblaciones de celulas enriquecidas en progenitores mieloides y/o linfoides y metodos de elaboracion y uso. |
EP0966523A1 (en) * | 1997-01-31 | 1999-12-29 | Hemosol Inc. | Method for the production of selected lymphocytes |
EP0983345A1 (en) * | 1997-05-21 | 2000-03-08 | THE UNITED STATES OF AMERICA, as represented by the Secretary of the Department of Health and Human Services | Methods and compositions for making dendritic cells from expanded populations of monocytes and for activating t cells |
AU753975B2 (en) * | 1997-12-04 | 2002-10-31 | Duke University | Methods of isolating and using CD7+CD34-Lin-hematopoietic cells |
FR2801056B1 (fr) * | 1999-11-12 | 2003-03-28 | Commissariat Energie Atomique | Proteine presente a la surface des cellules souches hematopoietiques de la lignee lymphoide et des cellules nk, et ses applications |
KR20020083634A (ko) * | 2001-04-27 | 2002-11-04 | 크레아젠 주식회사 | 수지상 세포를 포함하는 자가면역 질환의 치료용 약제학적조성물 및 이를 이용한 치료방법 |
US20050042751A1 (en) * | 2001-10-31 | 2005-02-24 | Michel Goldman | Generation and use of new types of dendritic cells |
JP4662776B2 (ja) * | 2002-12-04 | 2011-03-30 | ベイラー リサーチ インスティテュート | 抗原負荷した樹状細胞ワクチンを前駆体から発生させる迅速ワンステップ法 |
US20050032210A1 (en) * | 2003-03-18 | 2005-02-10 | Kirin Beer Kabushiki Kaisha | Method of preparing immuno-regulatory dendritic cells and the use thereof |
EP2335738A1 (en) * | 2003-04-01 | 2011-06-22 | United States Government as represented by the Department of Veteran's Affaires | Stem-cell, precursor cell or target cell-based treatment of wound healing |
WO2005085426A1 (ja) * | 2004-03-04 | 2005-09-15 | Tanabe Seiyaku Co., Ltd. | 無フィ−ダ−分化用培地及び霊長類動物胚性幹細胞からの無フィ−ダ−分化方法 |
CN101006172B (zh) * | 2004-06-24 | 2012-09-05 | 株式会社载体研究所 | 包含导入了rna病毒的树突状细胞的抗癌剂 |
CN101080487B (zh) * | 2004-10-07 | 2012-11-14 | 阿戈斯治疗公司 | 成熟树突细胞组合物及其培养方法 |
US20080166751A1 (en) * | 2005-02-23 | 2008-07-10 | Takayuki Asahara | Method of Analyzing Dynamics in the Differentiation of Vascular Endothelial Progenitor Cells |
KR20170086700A (ko) * | 2005-04-08 | 2017-07-26 | 아르고스 쎄라퓨틱스 인코포레이티드 | 수지상 세포 조성물 및 방법 |
AU2006252576B2 (en) * | 2005-06-01 | 2011-01-20 | Wisconsin Alumni Research Foundation | Method of forming dendritic cells from embryonic stem cells |
PL1941027T3 (pl) * | 2005-09-28 | 2015-01-30 | Ipd Therapeutics B V | Metody i środki do proliferacji komórek macierzystych oraz wytwarzania i ekspansji komórek progenitorowych, a także produkcji komórek efektorowych jako leków klinicznych |
KR102651433B1 (ko) | 2006-04-14 | 2024-03-25 | 아스텔라스 인스티튜트 포 리제너러티브 메디슨 | 혈관 콜로니 형성 세포 |
WO2008038279A2 (en) * | 2006-09-25 | 2008-04-03 | Kfir Nissan | Force transducer and method |
DE102006054041B3 (de) * | 2006-11-16 | 2008-05-08 | Pierburg Gmbh | Regelvorrichtung für eine Verbrennungskraftmaschine |
SI2203176T1 (sl) * | 2007-09-28 | 2015-04-30 | Anthrogenesis Corporation | Tumorska supresija z uporabo humanega perfuzata placente in intermediarnih naravnih celic ubijalk, pridobljenih iz humane placente |
SG10201610881UA (en) * | 2008-03-27 | 2017-03-30 | Asterias Biotherapeutics Inc | Differentiation of primate pluripotent stem cells to hematopoietic lineage cells |
ES2685969T3 (es) | 2008-05-06 | 2018-10-15 | Astellas Institute For Regenerative Medicine | Células formadoras de colonias hemangioblásticas y células hemangioblásticas no injertables |
KR20210003301A (ko) | 2008-05-06 | 2021-01-11 | 아스텔라스 인스티튜트 포 리제너러티브 메디슨 | 다능성 줄기세포로부터 유도된 탈핵 적혈구계 세포를 생산하는 방법 |
KR101720961B1 (ko) * | 2009-02-27 | 2017-03-29 | 셀룰러 다이내믹스 인터내셔널, 인코포레이티드 | 다능성 세포의 분화 |
BR112012014104A2 (pt) | 2009-12-04 | 2016-07-05 | Stem Cell & Regenerative Medicine International Inc | método de geração de células destrutoras naturais e células dendríticas de hemangioblastos embrionários humanos derivados de células-tronco |
-
2010
- 2010-12-01 BR BR112012014104A patent/BR112012014104A2/pt not_active IP Right Cessation
- 2010-12-01 CA CA2781969A patent/CA2781969A1/en not_active Abandoned
- 2010-12-01 CA CA3080368A patent/CA3080368A1/en not_active Abandoned
- 2010-12-01 EP EP10835074.5A patent/EP2507365A4/en not_active Withdrawn
- 2010-12-01 KR KR1020197003655A patent/KR20190016602A/ko not_active Application Discontinuation
- 2010-12-01 CA CA3095576A patent/CA3095576A1/en active Pending
- 2010-12-01 AU AU2010326027A patent/AU2010326027A1/en not_active Abandoned
- 2010-12-01 IN IN5053DEN2012 patent/IN2012DN05053A/en unknown
- 2010-12-01 JP JP2012542163A patent/JP6013187B2/ja active Active
- 2010-12-01 RU RU2012127811/10A patent/RU2012127811A/ru not_active Application Discontinuation
- 2010-12-01 KR KR1020217030824A patent/KR20210120136A/ko not_active Application Discontinuation
- 2010-12-01 WO PCT/US2010/058593 patent/WO2011068896A1/en active Application Filing
- 2010-12-01 KR KR1020127017370A patent/KR101947588B1/ko active IP Right Grant
- 2010-12-01 KR KR1020207018660A patent/KR20200083645A/ko not_active IP Right Cessation
- 2010-12-01 US US13/512,795 patent/US8822218B2/en active Active
- 2010-12-01 CN CN2010800629687A patent/CN102822332A/zh active Pending
-
2014
- 2014-08-21 US US14/465,533 patent/US20150140657A1/en not_active Abandoned
-
2016
- 2016-03-24 JP JP2016060101A patent/JP2016119910A/ja active Pending
-
2019
- 2019-02-12 JP JP2019022273A patent/JP7343984B2/ja active Active
- 2019-10-31 US US16/669,931 patent/US20200131475A1/en active Pending
-
2021
- 2021-11-19 JP JP2021188673A patent/JP2022027790A/ja active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070141703A1 (en) | 2003-11-19 | 2007-06-21 | Stanley Edouard G | Methods for producing blood products from pluripotent cells in cell culture |
WO2008056779A1 (fr) | 2006-11-09 | 2008-05-15 | Japan As Represented By The President Of International Medical Center Of Japan | Procédé destiné à la culture et au passage d'une cellule souche embryonnaire de primate, et procédé destiné à induire la différenciation de la cellule souche embryonnaire |
Non-Patent Citations (2)
Title |
---|
BLOOD, 2008年11月 1日,Vol.112, No.9, p.3601-3614 |
The Journal of Immunology, 2005, Vol.175, p.5095-5103 |
Also Published As
Publication number | Publication date |
---|---|
KR20120112511A (ko) | 2012-10-11 |
KR20190016602A (ko) | 2019-02-18 |
US20120282693A1 (en) | 2012-11-08 |
IN2012DN05053A (ja) | 2015-10-09 |
JP2016119910A (ja) | 2016-07-07 |
EP2507365A4 (en) | 2014-01-01 |
JP2022027790A (ja) | 2022-02-14 |
KR101947588B1 (ko) | 2019-02-14 |
US20150140657A1 (en) | 2015-05-21 |
JP6013187B2 (ja) | 2016-10-25 |
KR20210120136A (ko) | 2021-10-06 |
WO2011068896A1 (en) | 2011-06-09 |
JP2013512673A (ja) | 2013-04-18 |
BR112012014104A2 (pt) | 2016-07-05 |
RU2012127811A (ru) | 2014-01-10 |
US8822218B2 (en) | 2014-09-02 |
CN102822332A (zh) | 2012-12-12 |
KR20200083645A (ko) | 2020-07-08 |
JP2019083819A (ja) | 2019-06-06 |
CA3095576A1 (en) | 2011-06-09 |
US20200131475A1 (en) | 2020-04-30 |
EP2507365A1 (en) | 2012-10-10 |
CA3080368A1 (en) | 2011-06-09 |
AU2010326027A1 (en) | 2012-06-21 |
CA2781969A1 (en) | 2011-06-09 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7343984B2 (ja) | ヒト胚性幹細胞由来血管芽細胞からナチュラルキラー細胞および樹状細胞を生成する方法 | |
AU2020264375B2 (en) | Method for developing natural killer cells from stem cells | |
KR101923290B1 (ko) | 혈관 콜로니 형성 세포 및 비-생착성 혈관 세포 | |
JP2024026891A (ja) | 造血系統細胞を製造するための方法およびシステム | |
US20050221482A1 (en) | Methods and compositions for obtaining hematopoietic stem cells derived from embryonic stem cells and uses thereof | |
KR20110006705A (ko) | 다능성 줄기세포로부터 유도된 탈핵 적혈구계 세포를 생산하는 방법 | |
Cheng et al. | In Vitro Haematopoietic Differentiation from Pluripotent Stem Cells or by Direct Lineage Conversion: Current Advances and Challenges | |
Dahéron et al. | Generation of Definitive Engraftable Hematopoietic Stem Cells from Human Pluripotent Stem Cells | |
Palma et al. | Pluripotent Stem Cells-A Novel Source of Haematopoietic Cells for Transplantation and Transfusion Medicine |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20190314 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20190314 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20200127 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20200427 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20200917 |
|
A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20200928 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20200928 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20201216 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20210217 |
|
A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20210719 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20211119 |
|
C60 | Trial request (containing other claim documents, opposition documents) |
Free format text: JAPANESE INTERMEDIATE CODE: C60 Effective date: 20211119 |
|
C11 | Written invitation by the commissioner to file amendments |
Free format text: JAPANESE INTERMEDIATE CODE: C11 Effective date: 20211201 |
|
A911 | Transfer to examiner for re-examination before appeal (zenchi) |
Free format text: JAPANESE INTERMEDIATE CODE: A911 Effective date: 20211221 |
|
C21 | Notice of transfer of a case for reconsideration by examiners before appeal proceedings |
Free format text: JAPANESE INTERMEDIATE CODE: C21 Effective date: 20211222 |
|
A912 | Re-examination (zenchi) completed and case transferred to appeal board |
Free format text: JAPANESE INTERMEDIATE CODE: A912 Effective date: 20220128 |
|
C211 | Notice of termination of reconsideration by examiners before appeal proceedings |
Free format text: JAPANESE INTERMEDIATE CODE: C211 Effective date: 20220202 |
|
C22 | Notice of designation (change) of administrative judge |
Free format text: JAPANESE INTERMEDIATE CODE: C22 Effective date: 20221207 |
|
C13 | Notice of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: C13 Effective date: 20230201 |
|
C22 | Notice of designation (change) of administrative judge |
Free format text: JAPANESE INTERMEDIATE CODE: C22 Effective date: 20230406 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20230501 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20230601 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20230619 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20230901 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 7343984 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |