CN105985985B - Crispr技术编辑并用igf优化的异体间充质干细胞的制备方法及在治疗心梗中应用 - Google Patents
Crispr技术编辑并用igf优化的异体间充质干细胞的制备方法及在治疗心梗中应用 Download PDFInfo
- Publication number
- CN105985985B CN105985985B CN201610297972.0A CN201610297972A CN105985985B CN 105985985 B CN105985985 B CN 105985985B CN 201610297972 A CN201610297972 A CN 201610297972A CN 105985985 B CN105985985 B CN 105985985B
- Authority
- CN
- China
- Prior art keywords
- mesenchymal stem
- stem cells
- cells
- allogeneic mesenchymal
- allogeneic
- 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
- 210000002901 mesenchymal stem cell Anatomy 0.000 title claims abstract description 101
- 108091033409 CRISPR Proteins 0.000 title claims abstract description 74
- 230000000735 allogeneic effect Effects 0.000 title claims abstract description 49
- 238000005516 engineering process Methods 0.000 title claims abstract description 40
- 238000010354 CRISPR gene editing Methods 0.000 title claims abstract description 37
- 208000010125 myocardial infarction Diseases 0.000 title claims abstract description 32
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 title claims abstract description 31
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 102000013275 Somatomedins Human genes 0.000 title abstract description 6
- 238000000034 method Methods 0.000 claims abstract description 31
- 102000004218 Insulin-Like Growth Factor I Human genes 0.000 claims abstract description 25
- 239000002245 particle Substances 0.000 claims abstract description 19
- 238000002054 transplantation Methods 0.000 claims abstract description 19
- 230000001900 immune effect Effects 0.000 claims abstract description 16
- 230000002757 inflammatory effect Effects 0.000 claims abstract description 16
- 239000000427 antigen Substances 0.000 claims abstract description 13
- 102000036639 antigens Human genes 0.000 claims abstract description 13
- 108091007433 antigens Proteins 0.000 claims abstract description 13
- 206010061218 Inflammation Diseases 0.000 claims abstract description 10
- 230000004054 inflammatory process Effects 0.000 claims abstract description 10
- 239000003814 drug Substances 0.000 claims abstract description 9
- 238000004115 adherent culture Methods 0.000 claims abstract description 6
- 101710160107 Outer membrane protein A Proteins 0.000 claims abstract description 4
- 238000000432 density-gradient centrifugation Methods 0.000 claims abstract description 4
- 210000005087 mononuclear cell Anatomy 0.000 claims abstract description 4
- 210000004027 cell Anatomy 0.000 claims description 70
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 25
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 claims description 24
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 claims description 24
- 239000013612 plasmid Substances 0.000 claims description 19
- 108020004414 DNA Proteins 0.000 claims description 16
- 241000282414 Homo sapiens Species 0.000 claims description 14
- 239000003102 growth factor Substances 0.000 claims description 14
- 210000004700 fetal blood Anatomy 0.000 claims description 13
- 238000012258 culturing Methods 0.000 claims description 10
- 239000001963 growth medium Substances 0.000 claims description 10
- 102100031650 C-X-C chemokine receptor type 4 Human genes 0.000 claims description 9
- 101000922348 Homo sapiens C-X-C chemokine receptor type 4 Proteins 0.000 claims description 9
- 238000004806 packaging method and process Methods 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 8
- 108020005004 Guide RNA Proteins 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 7
- 230000001464 adherent effect Effects 0.000 claims description 6
- 239000006285 cell suspension Substances 0.000 claims description 6
- 230000001965 increasing effect Effects 0.000 claims description 6
- 239000000725 suspension Substances 0.000 claims description 6
- 230000006907 apoptotic process Effects 0.000 claims description 5
- 210000004369 blood Anatomy 0.000 claims description 5
- 239000008280 blood Substances 0.000 claims description 5
- 239000001506 calcium phosphate Substances 0.000 claims description 5
- 229910000389 calcium phosphate Inorganic materials 0.000 claims description 5
- 235000011010 calcium phosphates Nutrition 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 239000012528 membrane Substances 0.000 claims description 5
- 230000002107 myocardial effect Effects 0.000 claims description 5
- 238000012216 screening Methods 0.000 claims description 5
- 238000000926 separation method Methods 0.000 claims description 5
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 claims description 5
- 102000053602 DNA Human genes 0.000 claims description 4
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 4
- 230000004927 fusion Effects 0.000 claims description 4
- 238000013518 transcription Methods 0.000 claims description 4
- 230000035897 transcription Effects 0.000 claims description 4
- 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 claims description 3
- 241000713666 Lentivirus Species 0.000 claims description 3
- 206010028851 Necrosis Diseases 0.000 claims description 3
- 239000012228 culture supernatant Substances 0.000 claims description 3
- 210000003754 fetus Anatomy 0.000 claims description 3
- 229960002897 heparin Drugs 0.000 claims description 3
- 229920000669 heparin Polymers 0.000 claims description 3
- 210000004698 lymphocyte Anatomy 0.000 claims description 3
- 230000017074 necrotic cell death Effects 0.000 claims description 3
- 230000000865 phosphorylative effect Effects 0.000 claims description 3
- 230000000379 polymerizing effect Effects 0.000 claims description 3
- 208000034423 Delivery Diseases 0.000 claims description 2
- 102000004142 Trypsin Human genes 0.000 claims description 2
- 108090000631 Trypsin Proteins 0.000 claims description 2
- 230000003321 amplification Effects 0.000 claims description 2
- 230000010100 anticoagulation Effects 0.000 claims description 2
- 238000007865 diluting Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 208000028867 ischemia Diseases 0.000 claims description 2
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 2
- 230000002194 synthesizing effect Effects 0.000 claims description 2
- 239000012588 trypsin Substances 0.000 claims description 2
- 210000004271 bone marrow stromal cell Anatomy 0.000 claims 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims 1
- 229940079593 drug Drugs 0.000 claims 1
- 229910052760 oxygen Inorganic materials 0.000 claims 1
- 239000001301 oxygen Substances 0.000 claims 1
- 208000024172 Cardiovascular disease Diseases 0.000 abstract description 8
- 230000006909 anti-apoptosis Effects 0.000 abstract description 3
- 239000008055 phosphate buffer solution Substances 0.000 description 10
- 238000001514 detection method Methods 0.000 description 9
- 210000000130 stem cell Anatomy 0.000 description 9
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 8
- 230000007547 defect Effects 0.000 description 7
- 102000004889 Interleukin-6 Human genes 0.000 description 6
- 108090001005 Interleukin-6 Proteins 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 206010000891 acute myocardial infarction Diseases 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000001890 transfection Methods 0.000 description 5
- 230000007423 decrease Effects 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 238000011476 stem cell transplantation Methods 0.000 description 4
- 238000002560 therapeutic procedure Methods 0.000 description 4
- 108010074051 C-Reactive Protein Proteins 0.000 description 3
- 102100032752 C-reactive protein Human genes 0.000 description 3
- 230000002424 anti-apoptotic effect Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- 210000003954 umbilical cord Anatomy 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 108010040476 FITC-annexin A5 Proteins 0.000 description 2
- 206010021143 Hypoxia Diseases 0.000 description 2
- 208000037273 Pathologic Processes Diseases 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000010362 genome editing Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 2
- 230000001146 hypoxic effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000003472 neutralizing effect Effects 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 230000009054 pathological process Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000000630 rising effect Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 230000000087 stabilizing effect Effects 0.000 description 2
- 101150076800 B2M gene Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 238000010453 CRISPR/Cas method Methods 0.000 description 1
- 101150066398 CXCR4 gene Proteins 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 241000252212 Danio rerio Species 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 206010014824 Endotoxic shock Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 101000599951 Homo sapiens Insulin-like growth factor I Proteins 0.000 description 1
- 101000611183 Homo sapiens Tumor necrosis factor Proteins 0.000 description 1
- 241000725303 Human immunodeficiency virus Species 0.000 description 1
- 206010061216 Infarction Diseases 0.000 description 1
- 102100037852 Insulin-like growth factor I Human genes 0.000 description 1
- 108090001007 Interleukin-8 Proteins 0.000 description 1
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 1
- 241000282567 Macaca fascicularis Species 0.000 description 1
- 206010028594 Myocardial fibrosis Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 208000007014 Retinitis pigmentosa Diseases 0.000 description 1
- 206010040070 Septic Shock Diseases 0.000 description 1
- 108020004459 Small interfering RNA Proteins 0.000 description 1
- 102100040247 Tumor necrosis factor Human genes 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 1
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 238000011316 allogeneic transplantation Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 230000002429 anti-coagulating effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 210000004413 cardiac myocyte Anatomy 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 239000005482 chemotactic factor Substances 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000004217 heart function Effects 0.000 description 1
- 210000005003 heart tissue Anatomy 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 210000003917 human chromosome Anatomy 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 210000004263 induced pluripotent stem cell Anatomy 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000007574 infarction Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 239000005550 inflammation mediator Substances 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 229960002725 isoflurane Drugs 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000011133 mesenchymal stem cell therapy Methods 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 210000001778 pluripotent stem cell Anatomy 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000010837 poor prognosis Methods 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 210000004927 skin cell Anatomy 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 230000002861 ventricular Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000002689 xenotransplantation 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- 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
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem 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
- C12N2740/00—Reverse transcribing RNA viruses
- C12N2740/00011—Details
- C12N2740/10011—Retroviridae
- C12N2740/15011—Lentivirus, not HIV, e.g. FIV, SIV
- C12N2740/15041—Use of virus, viral particle or viral elements as a vector
- C12N2740/15043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Developmental Biology & Embryology (AREA)
- Epidemiology (AREA)
- Plant Pathology (AREA)
- Public Health (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Animal Behavior & Ethology (AREA)
- Molecular Biology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
Claims (9)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610297972.0A CN105985985B (zh) | 2016-05-06 | 2016-05-06 | Crispr技术编辑并用igf优化的异体间充质干细胞的制备方法及在治疗心梗中应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610297972.0A CN105985985B (zh) | 2016-05-06 | 2016-05-06 | Crispr技术编辑并用igf优化的异体间充质干细胞的制备方法及在治疗心梗中应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN105985985A CN105985985A (zh) | 2016-10-05 |
CN105985985B true CN105985985B (zh) | 2019-12-31 |
Family
ID=57044650
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610297972.0A Active CN105985985B (zh) | 2016-05-06 | 2016-05-06 | Crispr技术编辑并用igf优化的异体间充质干细胞的制备方法及在治疗心梗中应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN105985985B (zh) |
Families Citing this family (32)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3613852A3 (en) | 2011-07-22 | 2020-04-22 | President and Fellows of Harvard College | Evaluation and improvement of nuclease cleavage specificity |
US9163284B2 (en) | 2013-08-09 | 2015-10-20 | President And Fellows Of Harvard College | Methods for identifying a target site of a Cas9 nuclease |
US9359599B2 (en) | 2013-08-22 | 2016-06-07 | President And Fellows Of Harvard College | Engineered transcription activator-like effector (TALE) domains and uses thereof |
US9322037B2 (en) | 2013-09-06 | 2016-04-26 | President And Fellows Of Harvard College | Cas9-FokI fusion proteins and uses thereof |
US9737604B2 (en) | 2013-09-06 | 2017-08-22 | President And Fellows Of Harvard College | Use of cationic lipids to deliver CAS9 |
US9228207B2 (en) | 2013-09-06 | 2016-01-05 | President And Fellows Of Harvard College | Switchable gRNAs comprising aptamers |
US20150166982A1 (en) | 2013-12-12 | 2015-06-18 | President And Fellows Of Harvard College | Methods for correcting pi3k point mutations |
US10077453B2 (en) | 2014-07-30 | 2018-09-18 | President And Fellows Of Harvard College | CAS9 proteins including ligand-dependent inteins |
IL294014B1 (en) | 2015-10-23 | 2024-03-01 | Harvard College | Nucleobase editors and their uses |
AU2017306676B2 (en) | 2016-08-03 | 2024-02-22 | President And Fellows Of Harvard College | Adenosine nucleobase editors and uses thereof |
CA3033327A1 (en) | 2016-08-09 | 2018-02-15 | President And Fellows Of Harvard College | Programmable cas9-recombinase fusion proteins and uses thereof |
WO2018039438A1 (en) | 2016-08-24 | 2018-03-01 | President And Fellows Of Harvard College | Incorporation of unnatural amino acids into proteins using base editing |
CN110214180A (zh) | 2016-10-14 | 2019-09-06 | 哈佛大学的校长及成员们 | 核碱基编辑器的aav递送 |
WO2018119359A1 (en) | 2016-12-23 | 2018-06-28 | President And Fellows Of Harvard College | Editing of ccr5 receptor gene to protect against hiv infection |
US11898179B2 (en) | 2017-03-09 | 2024-02-13 | President And Fellows Of Harvard College | Suppression of pain by gene editing |
JP2020510439A (ja) | 2017-03-10 | 2020-04-09 | プレジデント アンド フェローズ オブ ハーバード カレッジ | シトシンからグアニンへの塩基編集因子 |
KR20190130613A (ko) | 2017-03-23 | 2019-11-22 | 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 | 핵산 프로그램가능한 dna 결합 단백질을 포함하는 핵염기 편집제 |
WO2018209320A1 (en) | 2017-05-12 | 2018-11-15 | President And Fellows Of Harvard College | Aptazyme-embedded guide rnas for use with crispr-cas9 in genome editing and transcriptional activation |
WO2019023680A1 (en) | 2017-07-28 | 2019-01-31 | President And Fellows Of Harvard College | METHODS AND COMPOSITIONS FOR EVOLUTION OF BASIC EDITORS USING PHAGE-ASSISTED CONTINUOUS EVOLUTION (PACE) |
US11319532B2 (en) | 2017-08-30 | 2022-05-03 | President And Fellows Of Harvard College | High efficiency base editors comprising Gam |
US11795443B2 (en) | 2017-10-16 | 2023-10-24 | The Broad Institute, Inc. | Uses of adenosine base editors |
CN110055223A (zh) * | 2018-01-19 | 2019-07-26 | 北京百奥赛图基因生物技术有限公司 | 一种B2m基因改造的免疫缺陷动物的制备方法及其应用 |
CN108441520B (zh) * | 2018-04-04 | 2020-07-31 | 苏州大学 | 利用CRISPR/Cas9系统构建的基因条件性敲除方法 |
CN108949691B (zh) * | 2018-07-17 | 2019-07-12 | 杭州观梓健康科技有限公司 | 一种制备可实时检测间充质干细胞衰老的细胞模型的方法 |
CN109082442B (zh) * | 2018-07-17 | 2019-09-20 | 杭州观梓健康科技有限公司 | 一种可解除免疫抑制并增强肿瘤靶向性杀伤的间充质干细胞的制备方法 |
CN109055373B (zh) * | 2018-07-17 | 2019-09-20 | 杭州观梓健康科技有限公司 | 一种在干细胞中进行基因定向敲入的方法 |
EP3942040A1 (en) | 2019-03-19 | 2022-01-26 | The Broad Institute, Inc. | Methods and compositions for editing nucleotide sequences |
KR20220108793A (ko) * | 2019-11-27 | 2022-08-03 | 더 보드 오브 리젠츠 오브 더 유니버시티 오브 텍사스 시스템 | Msc 세포의 대규모 조합된 car 형질 도입 및 crispr 유전자 편집 |
CN110951693A (zh) * | 2019-12-05 | 2020-04-03 | 北京兴元和济生命医学科技有限公司 | 一种利用CRISPR-Cas9系统对间充质干细胞定点敲入外源基因的方法 |
MX2022014008A (es) | 2020-05-08 | 2023-02-09 | Broad Inst Inc | Métodos y composiciones para la edición simultánea de ambas cadenas de una secuencia de nucleótidos de doble cadena objetivo. |
CN113999877B (zh) * | 2021-09-18 | 2024-04-19 | 东南大学 | 一种间充质干细胞药物的基因编辑方法及应用 |
CN117448379A (zh) * | 2023-12-22 | 2024-01-26 | 上海元戊医学技术有限公司 | iPSC来源的IL-10蛋白过表达MSC细胞株的构建方法及其应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102021143A (zh) * | 2010-11-24 | 2011-04-20 | 浙江大学 | 一种提高间充质干细胞迁移能力的预处理方法 |
CN102046193A (zh) * | 2008-05-27 | 2011-05-04 | 梅奥医学教育和研究基金会 | 利用细胞治疗心脏组织的组合物和方法 |
CN104046593A (zh) * | 2013-03-14 | 2014-09-17 | 浙江大学 | 一种低免疫原性的人细胞及其制备方法 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104726494B (zh) * | 2015-02-12 | 2018-10-23 | 中国人民解放军第二军医大学 | CRISPR-Cas9技术构建染色体易位干细胞及动物模型的方法 |
-
2016
- 2016-05-06 CN CN201610297972.0A patent/CN105985985B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102046193A (zh) * | 2008-05-27 | 2011-05-04 | 梅奥医学教育和研究基金会 | 利用细胞治疗心脏组织的组合物和方法 |
CN102021143A (zh) * | 2010-11-24 | 2011-04-20 | 浙江大学 | 一种提高间充质干细胞迁移能力的预处理方法 |
CN104046593A (zh) * | 2013-03-14 | 2014-09-17 | 浙江大学 | 一种低免疫原性的人细胞及其制备方法 |
Non-Patent Citations (1)
Title |
---|
Downregulation of B2M in Allogeneic MSC by CRISPR Technology Inhibits Immune Rejection and Enhances Cardiac Repair;Yangxing Li et al.;《Circulation》;20180609;第136卷 * |
Also Published As
Publication number | Publication date |
---|---|
CN105985985A (zh) | 2016-10-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105985985B (zh) | Crispr技术编辑并用igf优化的异体间充质干细胞的制备方法及在治疗心梗中应用 | |
KR101310578B1 (ko) | 심혈관 증상의 치료에 지방조직-유래 세포를 사용하는 방법 | |
ES2364689T3 (es) | Procedimiento de uso de células derivadas de tejido adiposo en el tratamiento de afecciones cardiovasculares. | |
JPWO2009133943A1 (ja) | 生体内機能的細胞の高効率採取法 | |
CN105769910B (zh) | 一种人羊膜间充质干细胞的应用 | |
CN114874982B (zh) | 一种增强脐带间充质干细胞分泌血管内皮生长因子的培养方法 | |
CN110075124B (zh) | AMSC-MALAT1-Exo用于制备治疗肝脏疾病的药物中的应用及其制备方法 | |
CN103861088A (zh) | 一种治疗原发性肝癌的干细胞制剂及其制备方法 | |
CN113197919A (zh) | 鹿茸干细胞外泌体在制备改善或治疗骨关节炎和延缓细胞衰老的产品中的应用 | |
CN111944748A (zh) | 一种用于治疗心肌梗死的高表达il-10的人脂肪间充质干细胞外泌体及其用途 | |
CN113559273B (zh) | 一种静脉注射用成体干细胞的预处理方法、成体干细胞注射液及应用 | |
CN106606512B (zh) | 一种用于治疗心肌梗死的混合细胞制剂及制备方法与应用 | |
CN106267413B (zh) | 艾滋病血浆净化器 | |
CN101182493A (zh) | 一种治疗放射性肠炎的转基因间充质干细胞及其制备方法 | |
CN106267425A (zh) | Aids免疫吸附治疗仪 | |
CN111825756A (zh) | 一种脐带间充质干细胞因子在nk细胞体外培养方面的应用 | |
CN110669792A (zh) | 基因修饰间充质干细胞、制备方法、应用及细胞治疗产品 | |
Lee et al. | Therapeutic effects of umbilical cord blood derived mesenchymal stem cell-conditioned medium on pulmonary arterial hypertension in rats | |
CN106267419B (zh) | Hiv免疫净化器 | |
CN106178163B (zh) | 艾滋病生物细胞免疫治疗仪 | |
CN105233303B (zh) | 一种通过基因改造血管内皮细胞促进创面愈合的方法及其应用 | |
CN103040867A (zh) | 骨髓间充质干细胞来源的微泡在制备治疗肾损伤药物中的应用 | |
CN112980784A (zh) | 一种人脐带间充质干细胞的分离培养方法及应用 | |
CN106267415B (zh) | 艾滋病净化治疗仪 | |
CN111925983A (zh) | 一种用于治疗心肌梗死的高表达il-10的人脂肪间充质干细胞外泌体的制备方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20231130 Address after: No. 105, Lane 399, Ruiyi Road, Zhoupu Town, Pudong New Area, Shanghai, 200120 Patentee after: Wang Xiaohan Patentee after: Lu Yang Address before: 215000 199 Ren Yan Road, Suzhou Industrial Park, Jiangsu Patentee before: SOOCHOW University |
|
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20240325 Address after: 200131, Building 8, 2nd Floor, Block A-02, No. 200 Newton Road, China (Shanghai) Pilot Free Trade Zone, Pudong New Area, Shanghai Patentee after: SHANGHAI XINRUITE BIOMEDICAL TECHNOLOGY Co.,Ltd. Country or region after: China Address before: No. 105, Lane 399, Ruiyi Road, Zhoupu Town, Pudong New Area, Shanghai, 200120 Patentee before: Wang Xiaohan Country or region before: China Patentee before: Lu Yang |