CN113943699B - 对抗高糖损伤的脐带间充质干细胞诱导液、方法及应用 - Google Patents
对抗高糖损伤的脐带间充质干细胞诱导液、方法及应用 Download PDFInfo
- Publication number
- CN113943699B CN113943699B CN202010689782.XA CN202010689782A CN113943699B CN 113943699 B CN113943699 B CN 113943699B CN 202010689782 A CN202010689782 A CN 202010689782A CN 113943699 B CN113943699 B CN 113943699B
- Authority
- CN
- China
- Prior art keywords
- mesenchymal stem
- umbilical cord
- sugar
- wound
- high sugar
- 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
- 208000027418 Wounds and injury Diseases 0.000 title claims abstract description 45
- 230000006698 induction Effects 0.000 title claims abstract description 35
- 210000002901 mesenchymal stem cell Anatomy 0.000 title claims abstract description 31
- 210000003954 umbilical cord Anatomy 0.000 title claims abstract description 28
- 230000006378 damage Effects 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 16
- 239000007788 liquid Substances 0.000 title claims abstract description 14
- 208000014674 injury Diseases 0.000 title claims abstract description 11
- 210000004027 cell Anatomy 0.000 claims abstract description 47
- 206010052428 Wound Diseases 0.000 claims abstract description 34
- 206010012601 diabetes mellitus Diseases 0.000 claims abstract description 23
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 claims abstract description 11
- 108010019598 Liraglutide Proteins 0.000 claims abstract description 8
- YSDQQAXHVYUZIW-QCIJIYAXSA-N Liraglutide Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCNC(=O)CC[C@H](NC(=O)CCCCCCCCCCCCCCC)C(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=C(O)C=C1 YSDQQAXHVYUZIW-QCIJIYAXSA-N 0.000 claims abstract description 8
- 229960002701 liraglutide Drugs 0.000 claims abstract description 8
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 claims description 12
- 230000004069 differentiation Effects 0.000 claims description 11
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 claims description 8
- 239000002609 medium Substances 0.000 claims description 8
- 229930182555 Penicillin Natural products 0.000 claims description 6
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 claims description 6
- 239000006166 lysate Substances 0.000 claims description 6
- 229940049954 penicillin Drugs 0.000 claims description 6
- 229960005322 streptomycin Drugs 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- 230000001939 inductive effect Effects 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 claims 1
- 241000700159 Rattus Species 0.000 abstract description 13
- 230000000694 effects Effects 0.000 abstract description 13
- 230000035876 healing Effects 0.000 abstract description 12
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 abstract description 10
- 239000003963 antioxidant agent Substances 0.000 abstract description 10
- 230000003078 antioxidant effect Effects 0.000 abstract description 10
- 238000000338 in vitro Methods 0.000 abstract description 9
- 230000029663 wound healing Effects 0.000 abstract description 8
- 210000004204 blood vessel Anatomy 0.000 abstract description 7
- 210000004369 blood Anatomy 0.000 abstract description 6
- 239000008280 blood Substances 0.000 abstract description 6
- 230000006870 function Effects 0.000 abstract description 6
- 230000001737 promoting effect Effects 0.000 abstract description 6
- 230000008439 repair process Effects 0.000 abstract description 5
- 238000011160 research Methods 0.000 abstract description 5
- 230000003064 anti-oxidating effect Effects 0.000 abstract description 4
- 210000005036 nerve Anatomy 0.000 abstract description 4
- 230000004060 metabolic process Effects 0.000 abstract description 3
- 210000002569 neuron Anatomy 0.000 abstract description 3
- 230000008929 regeneration Effects 0.000 abstract description 3
- 238000011069 regeneration method Methods 0.000 abstract description 3
- 206010072170 Skin wound Diseases 0.000 abstract description 2
- 230000002218 hypoglycaemic effect Effects 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- 210000000130 stem cell Anatomy 0.000 description 14
- 239000000243 solution Substances 0.000 description 12
- 239000001963 growth medium Substances 0.000 description 10
- 210000004271 bone marrow stromal cell Anatomy 0.000 description 9
- 108090000695 Cytokines Proteins 0.000 description 6
- 102000004127 Cytokines Human genes 0.000 description 6
- 238000004113 cell culture Methods 0.000 description 5
- 230000033115 angiogenesis Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 108010025020 Nerve Growth Factor Proteins 0.000 description 3
- 102000015336 Nerve Growth Factor Human genes 0.000 description 3
- 230000001464 adherent effect Effects 0.000 description 3
- 230000006907 apoptotic process Effects 0.000 description 3
- 210000000227 basophil cell of anterior lobe of hypophysis Anatomy 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 238000005520 cutting process Methods 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 229940053128 nerve growth factor Drugs 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 102000009024 Epidermal Growth Factor Human genes 0.000 description 2
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 2
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 2
- 101800000224 Glucagon-like peptide 1 Proteins 0.000 description 2
- 102400000322 Glucagon-like peptide 1 Human genes 0.000 description 2
- DTHNMHAUYICORS-KTKZVXAJSA-N Glucagon-like peptide 1 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 DTHNMHAUYICORS-KTKZVXAJSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 108010019160 Pancreatin Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 2
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 2
- 230000024245 cell differentiation Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 210000002950 fibroblast Anatomy 0.000 description 2
- 229940126864 fibroblast growth factor Drugs 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- 201000001421 hyperglycemia Diseases 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 210000002660 insulin-secreting cell Anatomy 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- 229940055695 pancreatin Drugs 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 230000037314 wound repair Effects 0.000 description 2
- 102100032528 C-type lectin domain family 11 member A Human genes 0.000 description 1
- 101710167766 C-type lectin domain family 11 member A Proteins 0.000 description 1
- 206010056340 Diabetic ulcer Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 101800003838 Epidermal growth factor Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 108090000367 Fibroblast growth factor 9 Proteins 0.000 description 1
- 102100037665 Fibroblast growth factor 9 Human genes 0.000 description 1
- 102100021866 Hepatocyte growth factor Human genes 0.000 description 1
- 101500028774 Homo sapiens Glucagon-like peptide 1 Proteins 0.000 description 1
- 101000898034 Homo sapiens Hepatocyte growth factor Proteins 0.000 description 1
- 101001076408 Homo sapiens Interleukin-6 Proteins 0.000 description 1
- 101000868152 Homo sapiens Son of sevenless homolog 1 Proteins 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 206010020710 Hyperphagia Diseases 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 208000028389 Nerve injury Diseases 0.000 description 1
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 description 1
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 206010040943 Skin Ulcer Diseases 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 239000012080 ambient air Substances 0.000 description 1
- 230000002424 anti-apoptotic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 210000003158 enteroendocrine cell Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 229940116977 epidermal growth factor Drugs 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- MGXWVYUBJRZYPE-YUGYIWNOSA-N incretin Chemical class C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1C=CC(O)=CC=1)[C@@H](C)O)[C@@H](C)CC)C1=CC=C(O)C=C1 MGXWVYUBJRZYPE-YUGYIWNOSA-N 0.000 description 1
- 239000000859 incretin Substances 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000302 ischemic effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 210000003041 ligament Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 230000011278 mitosis Effects 0.000 description 1
- 230000001483 mobilizing effect Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000008764 nerve damage Effects 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 210000004498 neuroglial cell Anatomy 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 210000000578 peripheral nerve Anatomy 0.000 description 1
- 206010036067 polydipsia Diseases 0.000 description 1
- 208000022530 polyphagia Diseases 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- GCYXWQUSHADNBF-AAEALURTSA-N preproglucagon 78-108 Chemical class C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 GCYXWQUSHADNBF-AAEALURTSA-N 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 231100000019 skin ulcer Toxicity 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 238000009168 stem cell therapy Methods 0.000 description 1
- 238000011476 stem cell transplantation Methods 0.000 description 1
- 238000009580 stem-cell therapy Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 238000011426 transformation method Methods 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 210000003606 umbilical vein Anatomy 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(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]([C@@H](C)O)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](CCCNC(N)=N)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)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)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=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
- 208000016261 weight loss Diseases 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
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/70—Undefined extracts
- C12N2500/80—Undefined extracts from animals
- C12N2500/84—Undefined extracts from animals from mammals
-
- 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
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
-
- 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
- C12N2501/30—Hormones
- C12N2501/335—Glucagon; Glucagon-like peptide [GLP]; Exendin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/999—Small molecules not provided for elsewhere
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Diabetes (AREA)
- Zoology (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Hematology (AREA)
- General Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Microbiology (AREA)
- Rheumatology (AREA)
- Emergency Medicine (AREA)
- Endocrinology (AREA)
- Obesity (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Dermatology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
本发明提供了对抗高糖损伤的脐带间充质干细胞诱导液、方法及应用。建立了一种模拟糖尿病患者体内的高糖环境诱导培养液，通过添加抗氧化剂GSH，降糖的利拉鲁肽和碱性成纤维细胞生长因子(bFGF)等物质，经过5～10天的体外高糖环境和抗氧化诱导，增加了间充质干细胞的在高糖环境下的细胞活性和功能，能够更有效的促进创面血管和神经的生成，达到移植治疗糖尿病创面的目的。研究证实，经本方法诱导培养的间充质干细胞可显著促进大鼠糖尿病创面皮肤创伤的愈合，具有更好的改善代谢、降低血糖、促进血管再生、促进神经细胞修复、伤口愈合的功能。为糖尿病创面的临床治疗选择提供了优良的种子细胞。
Description
技术领域
本发明属于干细胞培养技术领域，涉及对抗高糖损伤的脐带间充质干细胞诱导液、方法及应用。
背景技术
糖尿病创面愈合不良是糖尿病的严重并发症之一，也一直是是临床治疗的难题。创面愈合是一个非常复杂的过程，涉及到修复细胞、炎症细胞、血管生成和末梢神经受损等多方面的相互协调，需要通过细胞因子、抗氧化作用、能量代谢等细胞微环境进行非常精细的调控。任何因素的失衡均会导致愈合延迟、难以愈合或创面不愈合。
糖尿病患者体内环境复杂，长期的高血糖、高血脂、酮体增多、活性氧增加等，使自身的MSCs动员迁移及损伤修复的能力减弱，创面常常经久不愈。
间充质干细胞(Mesnchymal Stem Cells，MSCs)是一类具有极强的自我更新和多项分化的潜能的细胞，可向骨、软骨、肌肉、韧带、神经等多个方向分化。MSCs是通过分泌的多种细胞因子发挥作用的，而目前的研究发现，在不同的环境刺激下，MSCs的分泌谱会随之发生改变。MSCs局部应用可调控炎症、增加血管化和促进上皮化，被认为是创面最有前景的治疗方法之一。
近年研究显示干细胞疗法对糖尿病创面具有一定的治疗效果。其中脐带间充质干细胞(Umbilical Cord Mesnchymal Stem Cells，UC-MSCs)因来源广泛，免疫原性低，用于移植治疗糖尿病及其并发症研究已经日成趋势。研究显示间充质干细胞能够移动和归巢到受伤和缺血组织，并分泌生长因子以促进血管生成和细胞外基质重塑；同时干细胞还能分化成许多细胞类型，如成纤维细胞、脂肪细胞、上皮细胞等，通过分化作用促进血管生成、神经细胞修复促进糖尿病创面愈合。
上述研究通常采用的是常规的干细胞培养技术，虽然收获一定量的干细胞用于创面的修复，但是由于糖尿病患者复杂的高糖高氧化损伤的身体条件，常常存在治疗效果不理想、修复功效缓慢的现象。
发明内容
针对上述技术问题，本发明提供了对抗高糖损伤的脐带间充质干细胞诱导液、方法及应用。
为实现本发明的目的，本发明采用以下技术方案：
对抗高糖损伤的脐带间充质干细胞诱导液，包括：高糖DMEM培养基，所述高糖DMEM培养基中加有人源化血小板裂解物，GSH，利拉鲁肽，bFGF以及青霉素/链霉素双抗。
H-DMEM培养能够体外模拟糖尿病患者体内的高糖环境，将UC-MSCs在无血清的H-DMEM里培养，能够诱导MSCs耐受高糖环境，使MSCs移植后在糖尿病患者的高糖环境中更好的耐受存活。本诱导培养基选择H-DMEM作为基础培养基，增加UC-MSCs的输注效果。
作为优选，高糖DMEM培养基(H-DMEM)的葡萄糖浓度为4.5g/L。
人源化血小板裂解物，含有多种生长因子(血小板衍生生长因子，PDGF；血管内皮生长因子，VEGF；表皮生长因子，EGF等)和蛋白质，能够有效促进MSCs增殖，且无异源的动物成分，可替代胎牛血清为UC-MSCs提供营养支持，同时有效规避了胎牛血清可能带来的过敏和动物源性疾病风险。
作为优选，培养基中人源化血小板裂解物的浓度为5～10％。
GSH是细胞内含有巯基的抗氧化剂，通过直接参与抗氧化酶的酶促反应，起到抗氧化，抗凋亡作用。还能预防损伤，调整基因表达，调节细胞的代谢活性，参与信号转导系统，保护细胞免受氧化损伤，并使内环境处于稳定的还原态。高血糖环境刺激下，导致细胞氧化应激反应增强及抗氧化能力减弱。培养基中添加GSH可以保护间充质干细胞免受氧化应激损伤，保持MSCs的活性和功能，增加干细胞的输注效果。
作为优选，培养基中GSH的浓度为1～20mmol/L。
利拉鲁肽是一种人胰高糖素样肽-1(GLP-1)类似物，用于治疗糖尿病。GLP-1是肠道L细胞分泌的一种肠促胰素，可以通过促进胰岛β细胞增殖，抑制其凋亡来增加β细胞数量。既往研究已经证实，在干细胞分化过程中，GLP-1可以充当诱导分化剂促进干细胞向胰岛素分泌细胞的成熟，提高分化效率。本诱导培养基中添加利拉鲁肽可促进UC-MSC向胰岛素分泌细胞分化。
作为优选，培养基中利拉鲁肽的使用浓度范围是1～10nmol/L。
碱性成纤维细胞生长因子(bFGF)能促进纤维细胞的有丝分裂，同时可以刺激血管生成、在体内能维持神经元、神经胶质细胞的存活。能促进细胞的黏附、增值与分化。本诱导培养基中添加bFGF可有效促进创面中神经和血管的修复，更有效促进创面愈合。
作为优选，培养基中bFGF的使用浓度范围是10～30ng/mL。
作为优选，培养基中青霉素/链霉素双抗为100U/mL青霉素，100mg/L链霉素。
本发明还提供了对抗高糖损伤的脐带间充质干细胞诱导方法，包括将UC-MSCs的传代细胞接种到培养瓶中，加入上述诱导液，进行诱导分化培养。
作为优选，诱导时间为5～10天。
本发明还提供了通过上述诱导方法得到的脐带间充质干细胞在用于糖尿病创面的药物中的应用。
本发明建立了一种模拟糖尿病患者体内的高糖环境诱导培养液，通过添加抗氧化剂GSH，降糖的利拉鲁肽和碱性成纤维细胞生长因子(bFGF)等物质，经过5～10天的体外高糖环境和抗氧化诱导，增加了间充质干细胞的在高糖环境下的细胞活性和功能，能够更有效的促进创面血管和神经的生成，达到移植治疗糖尿病创面的目的。
本发明的有益效果如下：
(一)针对现有的诱导培养技术存在细胞活性低，诱导转化方法复杂等缺陷，本发明设计了一种体外高糖和抗氧化诱导间充质干细胞的方法。
(二)本发明提供的诱导液用于体外诱导UC-MSCs耐受高糖环境，并且抵抗由于高糖诱导发生的氧化应激，能够减少细胞凋亡损伤。
(三)本发明建立的对抗高糖损伤诱导液诱导的IUC-MSCs能够加速大鼠糖尿病溃疡皮肤创伤的愈合，具有更好的改善代谢、降低血糖、促进血管再生、伤口愈合的功能。为糖尿病创面的临床治疗选择提供了优良的种子细胞。
(四)本发明建立的对抗高糖损伤诱导液诱导的IUC-MSCs相比常规UC-MSCs能分泌更高的HGF、FGF-9、NGF等细胞因子。
附图说明
图1为本发明实施例诱导分化过程中细胞形态学观察图，A.IUC-MSCs细胞；B.常规UC-MSCs细胞；C.单纯高糖培养液培养的UC-MSCs细胞。
图2为本发明实施例体外分泌细胞因子鉴定结果。
图3为本发明实施例三组大鼠创面愈合趋势折线图，A.PBS对照组；B.UC-MSCs组；C.IUC-MSCs组。
具体实施方式
下面对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
实施例
1.UC-MSCs体外分离培养
(1)脐带来源间充质干细胞的分离和原代细胞培养
1)无菌条件下，将脐带(脐带组织采集自足月剖宫产的健康产妇，采集之前与产妇及其家属均签署知情同意书，方案经医院医学伦理会批准。)用含青霉素/链霉素双抗的生理盐水反复冲洗，洗去脐带残留血液，将脐带剪成3.0-4.0cm的小段，每段均沿脐静脉腔剪开，平铺后剔除静脉和动脉，取出血管之间、血管与外膜之间的胶状物—华通胶，用小剪刀将华通胶剪切成约1mm3小块，接种于T75培养瓶内，使脐带华通胶均匀分布瓶底。置于二氧化碳培养箱内培养，培养箱内温度为37℃，CO2浓度为5％，贴壁4～18hr后加入15ml DM-PL干细胞培养液。进行原代细胞的爬出。
DM-PL干细胞培养液的配制：DMEM/F12基础培养液加入体积分数为5％人源化血小板裂解物。
2)5d后首次全量换液，弃去未贴壁细胞，每3～4d换液一次。倒置相差显微镜观察贴壁细胞，待细胞游出后，待细胞长至75％～85％融合时，采用0.25％胰蛋白酶于37℃消化贴壁细胞不超过5分钟。加入完全培养液终止消化，收集细胞悬液置50ml离心管，离心，1200rpm，5min，弃去含胰酶上清液，生理盐水清洗两次，弃上清，得到原代人UC-MSCs。
(2)细胞传代培养
将获得的原代细胞置于新的培养液中进行传代培养，倒置显微镜观察贴壁细胞融合达75％～85％，胰酶消化，生理盐水清洗2次，得到P1细胞；加入新的培养液按照1:3～1:4的比例重新进行接种，进行传代培养，并使用同样方法重复进行P2和P3代人脐带间充质干细胞的培养。培养温度为37℃，环境空气为5％CO2。
2.UC-MSCs体外诱导培养
将P3代UC-MSCs接种到新的T75培养瓶中，加入高糖抗氧化损伤培养液，进行诱导分化培养。常规培养基培养的细胞和单纯高糖抗氧化损伤培养液培养的细胞为对照细胞。诱导时间为9天。
常规培养基成分为DMEM培养液中添加10％FBS；
高糖抗氧化损伤培养液成分为高糖DMEM培养基(H-DMEM，4.5g葡萄糖/L)中添加5％人源化血小板裂解物，10mmol/L GSH，5nmol/L利拉鲁肽，100U/mL青霉素，100mg/L链霉素，20ng/mL bFGF。
3.诱导分化过程中细胞形态学观察
本发明诱导获得的IUC-MSCs细胞(A图)可稳定贴壁生长，所获得的细胞生物学特征同常规培养方法(B图)得到的细胞特征相似，显微镜下细胞为典型的梭形漩涡状两者都呈现梭行，为正常干细胞形态，两者没有大的差别。
单独用高糖培养液(C图)培养的UC-MSCs细胞，细胞生长缓慢，部分细胞变圆，胞核较大，细胞界限清晰。后期基本停止生长，出现凋亡状态。
4.UC-MSCs体外分泌细胞因子鉴定
实验分为2组：①UC-MSCs：脐带间充质干细胞组；②IUC-MSCs，诱导后脐带间充质干细胞组。分别取两组细胞上清液进行干细胞生长因子(HGF)、成纤维细胞生长因子(FGF)、神经生长因子(NGF)的检测。如图2可见，经IUC-MSCs组间充质干细胞分泌的组织创伤修复相关细胞因子显著高于非诱导组。且具有统计学意义，***，P<0.001，**P<0.01。
5.UC-MSCs治疗糖尿病创面
(1)糖尿病创面试验模型建立
Wistar雄性大鼠(6-8周龄，160-180g重)适应性饲养1周后，腹腔注射链脲佐菌素(streptozotocin，STZ)制作大鼠糖尿病模型，注射3天后，监测大鼠的血糖、体重、饮水量和尿量，待出现血糖≥16.7mmol/L及多饮、多尿、多食且体重减轻等糖尿病症状后，用打孔器在大鼠背部制作6mm×6mm的圆形全层皮肤缺损创面，建立大鼠糖尿病创面模型。
(2)干细胞准备
将诱导培养好的UC-MSCs细胞终止培养，生理盐水冲洗2遍，0.25％胰蛋白消化，进行细胞活性检测。细胞活性90％以上的细胞进行计数，PBS清洗两遍，将不同数量细胞分别溶于1ml PBS溶液，混匀备用。每毫升细胞悬液所含细胞数量为0.5～1.5×107。
(3)干细胞移植治疗
1)造模成功后，大鼠随机分成3组：A组为糖尿病创面对照组(PBS对照组)；B组为常规脐带间充质干细胞组(UC-MSCs组)；C组为高糖抗氧化诱导培养脐带间充质干细胞组(IUC-MSCs组)。每组15只大鼠。
2)用微量进样器抽取细胞悬液UC-MSCs及IUC-MSCs后沿创面边缘皮下注射，每个创面选择8个注射点，每点注射10μl。对照组抽取PBS沿创面边缘皮下注射，每个创面选择8个注射点均匀注射，每点注射10μl。
3)对各组大鼠分别在创面形成术后当天(0天)、第3天、第7天、第11天、第14天的创面愈合情况跟踪检测。计算不同时间点的创面愈合率，公式如下：
愈合率＝(原始创面面积－剩余创面面积)/原始创面面积×100％
(4)实验结果
第14天IUC-MSCs组大鼠创面的愈合率时间为79％，与UC-MSCs组相比无差异。PBS组愈合率较低48％，与各组相比均有显著差异(P<0.05)。与PBS对照组相比，局部注射脐带间充质干细胞能明显加快皮肤溃疡面积的愈合。两组干细胞组相比，用高糖抗氧化损伤诱导过的UC-MSCs具有更好的促进糖尿病创面皮肤愈合效果，表明诱导的IUC-MSCs具有比普通间充质干细胞更好的改善代谢、降低血糖、促进血管再生、伤口愈合的功能。
Claims (5)
1.对抗高糖损伤的脐带间充质干细胞的诱导方法，其特征在于，所述方法包括将脐带间充质干细胞的传代细胞接种到培养瓶中，加入对抗高糖损伤的脐带间充质干细胞诱导液，进行诱导分化培养获得对抗高糖损伤的脐带间充质干细胞；所述对抗高糖损伤的脐带间充质干细胞诱导液为添加有5％人源化血小板裂解物，10mmol/L GSH，5nmol/L利拉鲁肽，100U/mL青霉素，100mg/L链霉素和20ng/mL bFGF的高糖DMEM培养基。
2.根据权利要求1所述的诱导方法，其中所述高糖DMEM培养基中葡萄糖浓度为4.5g/L。
3.根据权利要求1或2所述的诱导方法，其特征在于，所述诱导分化培养为5～10天。
4.根据权利要求1或2所述的诱导方法，其特征在于，所述诱导分化培养为9天。
5.使用根据权利要求1-4任一项所述诱导方法获得的对抗高糖损伤的脐带间充质干细胞在制备用于治疗糖尿病创面的药物中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010689782.XA CN113943699B (zh) | 2020-07-17 | 2020-07-17 | 对抗高糖损伤的脐带间充质干细胞诱导液、方法及应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010689782.XA CN113943699B (zh) | 2020-07-17 | 2020-07-17 | 对抗高糖损伤的脐带间充质干细胞诱导液、方法及应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113943699A CN113943699A (zh) | 2022-01-18 |
| CN113943699B true CN113943699B (zh) | 2023-10-27 |
Family
ID=79326999
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010689782.XA Active CN113943699B (zh) | 2020-07-17 | 2020-07-17 | 对抗高糖损伤的脐带间充质干细胞诱导液、方法及应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113943699B (zh) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114480252B (zh) * | 2022-01-24 | 2023-04-25 | 四川大学华西医院 | 功能增强型内皮细胞的驯化方法及驯化得到的细胞制剂 |
| CN117138028B (zh) * | 2023-10-30 | 2024-02-13 | 北京岷德生物科技有限公司 | 一种含有表皮干细胞因子的组合物及其在糖尿病足修复中的应用 |
| CN117323411B (zh) * | 2023-11-02 | 2024-08-06 | 徐州医科大学附属医院 | 破伤风类毒素在脐带干细胞损伤治疗中的应用 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103756954A (zh) * | 2014-01-22 | 2014-04-30 | 青岛中天干细胞工程有限公司 | 一种将人脂肪间充质干细胞诱导成胰岛素分泌细胞的诱导剂、诱导培养基及方法 |
| KR20150115304A (ko) * | 2014-04-03 | 2015-10-14 | 주식회사 녹십자랩셀 | 중간엽 줄기세포 스페로이드로부터 분화된 인슐린 분비세포의 제조방법 및 그의 용도 |
| CN105596372A (zh) * | 2015-12-15 | 2016-05-25 | 天津卫硕生物科技有限公司 | 一种利用自体骨髓间充质干细胞治疗各种严重皮肤损伤的方法 |
| CN110115755A (zh) * | 2019-01-23 | 2019-08-13 | 温州医科大学 | 谷胱甘肽的医药新用途 |
-
2020
- 2020-07-17 CN CN202010689782.XA patent/CN113943699B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103756954A (zh) * | 2014-01-22 | 2014-04-30 | 青岛中天干细胞工程有限公司 | 一种将人脂肪间充质干细胞诱导成胰岛素分泌细胞的诱导剂、诱导培养基及方法 |
| KR20150115304A (ko) * | 2014-04-03 | 2015-10-14 | 주식회사 녹십자랩셀 | 중간엽 줄기세포 스페로이드로부터 분화된 인슐린 분비세포의 제조방법 및 그의 용도 |
| CN105596372A (zh) * | 2015-12-15 | 2016-05-25 | 天津卫硕生物科技有限公司 | 一种利用自体骨髓间充质干细胞治疗各种严重皮肤损伤的方法 |
| CN110115755A (zh) * | 2019-01-23 | 2019-08-13 | 温州医科大学 | 谷胱甘肽的医药新用途 |
Non-Patent Citations (4)
| Title |
|---|
| Liraglutide Up-regulation Thioredoxin Attenuated Müller Cells Apoptosis in High Glucose by Regulating Oxidative Stress and Endoplasmic Reticulum Stress;Xiang Ren等;《CURRENT EYE RESEARCH》;第1-2页 * |
| 利拉鲁肽对缺氧和高糖诱导的心肌细胞氧化应激损伤的保护作用;曾慧琳等;《中国药师》;第21卷(第05期);第783-786, 791页 * |
| 趋化因子-8对高糖诱导人间充质干细胞损伤的保护作用;沈雷等;《中国老年学杂志》;第38卷(第04期);第918-920页 * |
| 高糖诱导肾小管上皮细胞自噬激活以及GLP-1类似物利拉鲁肽对其调节作用的研究;赵昕等;《中国博士学位论文全文数据库 医药卫生科技辑》(第3期);第38、53页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113943699A (zh) | 2022-01-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113943699B (zh) | 对抗高糖损伤的脐带间充质干细胞诱导液、方法及应用 | |
| Auger et al. | Skin substitutes and wound healing | |
| KR101639988B1 (ko) | 고농도의 세포성장인자를 포함하는 중간엽 줄기세포 배양액의 제조방법 및 이로부터 얻어진 조성물 | |
| WO2016168950A1 (zh) | 一种唾液腺类器官和类腺泡的体外构建方法 | |
| CN104603263A (zh) | 高浓度干细胞的生产方法 | |
| CN113244272B (zh) | 用于改善卵巢早衰的组合物及其制备方法和应用 | |
| 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 | |
| CN110713984B (zh) | 诱导人间充质干细胞生成功能内皮细胞的方法 | |
| CN106190951A (zh) | 脂肪间充质干细胞诱导成肝脏样细胞的方法 | |
| CN105820998A (zh) | 临床回输级细胞治疗用人脂肪干细胞ADSCs分离提取培养方法 | |
| AU2007265862A1 (en) | Soft tissue filler composition comprising autologous dermis-derived cell culture product and hyaluronic acid | |
| US20100022003A1 (en) | Therapeutic cell medicine comprising skin tissue derived stem cell | |
| WO2024113917A1 (zh) | 毛囊细胞构成的多层组织工程皮肤及其制备方法与应用 | |
| CN112538513B (zh) | 细胞外基质mb生物蛋白及其制备试剂盒与方法 | |
| CN109929800A (zh) | 一种干细胞分泌因子的获取与纯化方法 | |
| CN110898078A (zh) | 一种间充质干细胞分泌因子的制备及应用 | |
| Julianto et al. | Topical delivery of mesenchymal stem cells “secretomes” in wound repair | |
| CN105013014A (zh) | 一种无细胞基质生物材料的制备方法和用途 | |
| CN114164171B (zh) | 一种人脐带间充质干细胞培养基、注射液、制备方法与其在制备治疗脑卒中的药物中的应用 | |
| CN111826343A (zh) | 一种增强诱导软骨分化的细胞培养液、方法及应用 | |
| CN105456293A (zh) | 一种用于治疗糖尿病的干细胞制剂及其制备方法 | |
| CN111172103A (zh) | 一种当归提取物刺激的干细胞外泌体的制备方法及其应用 | |
| CN110904049A (zh) | 一种富含vegf和fgf的干细胞上清液的制备方法及其应用 | |
| KR20150138700A (ko) | 건 또는 인대 손상 치유를 위한 자가 및 동종의 지방유래 중간엽줄기세포 조성물 및 이의 제조방법 | |
| CN111097041A (zh) | 一种利用间充质干细胞培养基联合纤维连接蛋白进行妊娠纹修复的混合液制备方法 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |