CN110403959B - 间充质干细胞外泌体制剂及其应用 - Google Patents
间充质干细胞外泌体制剂及其应用 Download PDFInfo
- Publication number
- CN110403959B CN110403959B CN201910867109.8A CN201910867109A CN110403959B CN 110403959 B CN110403959 B CN 110403959B CN 201910867109 A CN201910867109 A CN 201910867109A CN 110403959 B CN110403959 B CN 110403959B
- Authority
- CN
- China
- Prior art keywords
- mesenchymal stem
- exosome
- formulation
- osteoporosis
- stem 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
- 210000001808 exosome Anatomy 0.000 title claims abstract description 71
- 210000002901 mesenchymal stem cell Anatomy 0.000 title claims abstract description 50
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 208000001132 Osteoporosis Diseases 0.000 claims abstract description 39
- 239000003814 drug Substances 0.000 claims abstract description 7
- 239000003937 drug carrier Substances 0.000 claims abstract description 5
- 210000001185 bone marrow Anatomy 0.000 claims description 18
- 238000009472 formulation Methods 0.000 claims description 17
- 239000000203 mixture Substances 0.000 claims description 17
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 claims description 16
- 229960001138 acetylsalicylic acid Drugs 0.000 claims description 16
- 238000011282 treatment Methods 0.000 claims description 16
- 239000007924 injection Substances 0.000 claims description 4
- 238000002347 injection Methods 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 239000002775 capsule Substances 0.000 claims description 2
- 239000002674 ointment Substances 0.000 claims description 2
- 239000008363 phosphate buffer Substances 0.000 claims description 2
- 239000002504 physiological saline solution Substances 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 230000037182 bone density Effects 0.000 abstract description 8
- 235000015872 dietary supplement Nutrition 0.000 abstract description 3
- 239000000047 product Substances 0.000 abstract description 2
- 241000699670 Mus sp. Species 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 239000000243 solution Substances 0.000 description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 210000000988 bone and bone Anatomy 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 230000002293 adipogenic effect Effects 0.000 description 4
- 210000002798 bone marrow cell Anatomy 0.000 description 4
- 238000002955 isolation Methods 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 229950004398 broxuridine Drugs 0.000 description 3
- 239000002458 cell surface marker Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 230000002188 osteogenic effect Effects 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- WOVKYSAHUYNSMH-RRKCRQDMSA-N 5-bromodeoxyuridine Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C(Br)=C1 WOVKYSAHUYNSMH-RRKCRQDMSA-N 0.000 description 2
- 102000007372 Ataxin-1 Human genes 0.000 description 2
- 108010032963 Ataxin-1 Proteins 0.000 description 2
- WOVKYSAHUYNSMH-UHFFFAOYSA-N BROMODEOXYURIDINE Natural products C1C(O)C(CO)OC1N1C(=O)NC(=O)C(Br)=C1 WOVKYSAHUYNSMH-UHFFFAOYSA-N 0.000 description 2
- 229940122361 Bisphosphonate Drugs 0.000 description 2
- 208000010392 Bone Fractures Diseases 0.000 description 2
- 208000006386 Bone Resorption Diseases 0.000 description 2
- 102000055006 Calcitonin Human genes 0.000 description 2
- 108060001064 Calcitonin Proteins 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- 102100037241 Endoglin Human genes 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 206010017076 Fracture Diseases 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 description 2
- 101000881679 Homo sapiens Endoglin Proteins 0.000 description 2
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 description 2
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 2
- 101000800116 Homo sapiens Thy-1 membrane glycoprotein Proteins 0.000 description 2
- 102000003992 Peroxidases Human genes 0.000 description 2
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 2
- 208000009415 Spinocerebellar Ataxias Diseases 0.000 description 2
- 102100033523 Thy-1 membrane glycoprotein Human genes 0.000 description 2
- 229930003316 Vitamin D Natural products 0.000 description 2
- QYSXJUFSXHHAJI-XFEUOLMDSA-N Vitamin D3 Natural products C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C/C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-XFEUOLMDSA-N 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 150000004663 bisphosphonates Chemical class 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 230000024279 bone resorption Effects 0.000 description 2
- 229960004015 calcitonin Drugs 0.000 description 2
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000002648 combination therapy Methods 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 238000011010 flushing procedure Methods 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- CGIGDMFJXJATDK-UHFFFAOYSA-N indomethacin Chemical compound CC1=C(CC(O)=O)C2=CC(OC)=CC=C2N1C(=O)C1=CC=C(Cl)C=C1 CGIGDMFJXJATDK-UHFFFAOYSA-N 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 2
- 235000008696 isoflavones Nutrition 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 201000003624 spinocerebellar ataxia type 1 Diseases 0.000 description 2
- 210000000130 stem cell Anatomy 0.000 description 2
- 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 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 210000002303 tibia Anatomy 0.000 description 2
- 210000000689 upper leg Anatomy 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 239000011710 vitamin D Substances 0.000 description 2
- 235000019166 vitamin D Nutrition 0.000 description 2
- 150000003710 vitamin D derivatives Chemical class 0.000 description 2
- 229940046008 vitamin d Drugs 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 125000005273 2-acetoxybenzoic acid group Chemical group 0.000 description 1
- APIXJSLKIYYUKG-UHFFFAOYSA-N 3 Isobutyl 1 methylxanthine Chemical compound O=C1N(C)C(=O)N(CC(C)C)C2=C1N=CN2 APIXJSLKIYYUKG-UHFFFAOYSA-N 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 238000011735 C3H mouse Methods 0.000 description 1
- 102100027221 CD81 antigen Human genes 0.000 description 1
- 102000015775 Core Binding Factor Alpha 1 Subunit Human genes 0.000 description 1
- 108010024682 Core Binding Factor Alpha 1 Subunit Proteins 0.000 description 1
- 101000914479 Homo sapiens CD81 antigen Proteins 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- NPGIHFRTRXVWOY-UHFFFAOYSA-N Oil red O Chemical compound Cc1ccc(C)c(c1)N=Nc1cc(C)c(cc1C)N=Nc1c(O)ccc2ccccc12 NPGIHFRTRXVWOY-UHFFFAOYSA-N 0.000 description 1
- 206010031264 Osteonecrosis Diseases 0.000 description 1
- 102000003982 Parathyroid hormone Human genes 0.000 description 1
- 108090000445 Parathyroid hormone Proteins 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- 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 description 1
- 108090000029 Peroxisome Proliferator-Activated Receptors Proteins 0.000 description 1
- 102100038831 Peroxisome proliferator-activated receptor alpha Human genes 0.000 description 1
- 108010090804 Streptavidin Proteins 0.000 description 1
- 238000003917 TEM image Methods 0.000 description 1
- 206010064390 Tumour invasion Diseases 0.000 description 1
- 230000009815 adipogenic differentiation Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- RGCKGOZRHPZPFP-UHFFFAOYSA-N alizarin Chemical compound C1=CC=C2C(=O)C3=C(O)C(O)=CC=C3C(=O)C2=C1 RGCKGOZRHPZPFP-UHFFFAOYSA-N 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 230000037118 bone strength Effects 0.000 description 1
- 230000009400 cancer invasion Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000011284 combination treatment Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 229960003957 dexamethasone Drugs 0.000 description 1
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000001177 diphosphate Substances 0.000 description 1
- XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical compound [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 description 1
- 235000011180 diphosphates Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000004821 effect on bone Effects 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 238000009164 estrogen replacement therapy Methods 0.000 description 1
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethyl mercaptane Natural products CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 210000003195 fascia Anatomy 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 150000002222 fluorine compounds Chemical class 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 210000003958 hematopoietic stem cell Anatomy 0.000 description 1
- 238000001794 hormone therapy Methods 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 229960000905 indomethacin Drugs 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- GOMNOOKGLZYEJT-UHFFFAOYSA-N isoflavone Chemical compound C=1OC2=CC=CC=C2C(=O)C=1C1=CC=CC=C1 GOMNOOKGLZYEJT-UHFFFAOYSA-N 0.000 description 1
- 150000002515 isoflavone derivatives Chemical class 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 239000002417 nutraceutical Substances 0.000 description 1
- 235000021436 nutraceutical agent Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000000879 optical micrograph Methods 0.000 description 1
- 230000009818 osteogenic differentiation Effects 0.000 description 1
- 230000002611 ovarian Effects 0.000 description 1
- 239000000199 parathyroid hormone Substances 0.000 description 1
- 229960001319 parathyroid hormone Drugs 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000000333 selective estrogen receptor modulator Substances 0.000 description 1
- 229940095743 selective estrogen receptor modulator Drugs 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/60—Salicylic acid; Derivatives thereof
- A61K31/612—Salicylic acid; Derivatives thereof having the hydroxy group in position 2 esterified, e.g. salicylsulfuric acid
- A61K31/616—Salicylic acid; Derivatives thereof having the hydroxy group in position 2 esterified, e.g. salicylsulfuric acid by carboxylic acids, e.g. acetylsalicylic acid
-
- 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
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Physical Education & Sports Medicine (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Immunology (AREA)
- Epidemiology (AREA)
- Rheumatology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Polymers & Plastics (AREA)
- Zoology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Hematology (AREA)
- Biotechnology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Virology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明提供了一种间充质干细胞外泌体制剂,其中,所述间充质干细胞外泌体制剂包括间充质干细胞来源的外泌体和药学上可接受的载体。本发明还提供了间充质干细胞外泌体或间充质干细胞外泌体制剂在制备用于治疗或预防骨质疏松症或者增加骨密度的药物、保健品或营养补充剂中的应用。
Description
技术领域
本发明总体涉及生物医药领域,具体地涉及一种间充质干细胞外泌体制剂及其用途。
背景技术
骨质疏松症是由多种原因引起的一种全身性骨骼疾病,其以骨强度降低和骨折危险性增加为特征。随着人类寿命的不断延长,老龄化问题也日趋严峻,骨质疏松症及其引起的骨折风险严重威胁着中老年人的健康,日益成为全球关注的焦点问题。我国对骨质疏松症进行抽样流行病学研究的结果表明:在40岁以上人群中,骨质疏松症及低骨量的患病率女性分别为19.9%和32.4%,而男性分别为11.5%和45.8%。60岁以上人群的骨质疏松症及低骨量患病率:女性分别为28.6%和13.8%,男性分别为15%和12.7%。这些数据显示骨质疏松症在中老年人群的高发生率,因此预防和治疗骨质疏松症对于维持中老年人群的健康显得尤为重要。
我国目前是世界人口大国,亦是老年人口绝对数量最多的国家,只有及早采取且制定合理防治骨质疏松症的措施和方法,才能在全国范围内有效地防治骨质疏松症。目前,骨质疏松症的预防方法主要有加强运动和营养,其在一定程度上可以预防骨质疏松症的发生发展。骨质疏松症的治疗方法主要包括雌激素代替疗法、降钙素、选择性雌激素受体调节剂以及二磷酸盐疗法,这些药物可以阻止骨吸收但对骨形成的作用特别小。具体地,用于治疗和阻止骨质疏松症发展的药物可分为两大类,第一类为抑制骨吸收药,包括钙剂、维生素D及活性维生素D、降钙素、二磷酸盐、雌激素以及异黄酮等;第二类为促进骨形成药,包括氟化物、合成类固醇、甲状旁腺激素以及异黄酮等。其中激素治疗及二磷酸盐的治疗容易引起易发感染及骨坏死等多种并发症。因此,寻找一种新的、高效且不具有或具有极少不良副作用的治疗和预防骨质疏松症的药物迫在眉睫。
外泌体是多种细胞在正常及病理状态下均可分泌到细胞外的包含复杂RNA和蛋白质的小膜泡。研究发现,外泌体可通过其携带的蛋白质、核酸、脂类等来调节受体细胞的生物学活性。另外,已有研究发现外泌体具有多种功能,例如其可参与到机体的免疫应答、细胞迁移、抗原提呈、细胞分化、肿瘤侵袭等生物过程中。
发明内容
本发明提供了一种间充质干细胞外泌体制剂及其应用,从而为骨质疏松症的防治提供一种新的途径。
一方面,本发明提供了一种用于治疗或预防骨质疏松症的间充质干细胞外泌体制剂,其包括间充质干细胞来源的外泌体和药学上可接受的载体。
进一步地,所述外泌体制剂还包含乙酰水杨酸。
进一步地,所述外泌体和所述乙酰水杨酸的质量比为1:1至3:1。
进一步地,相对于所述外泌体制剂的总体积,所述外泌体制剂中的外泌体含量为500μg/ml至2500μg/ml,优选750μg/ml至1500μg/ml,最优选1000μg/ml。
进一步地,所述制剂包含100μg至500μg、优选150μg至300μg、最优选200μg的外泌体。
进一步地,所述外泌体的直径为40nm至250nm、优选地40nm至200nm。
进一步地,所述间充质干细胞选自骨髓间充质干细胞、口腔干细胞、脂肪来源的间充质细胞、人脐带血来源的间充质干细胞和人胚胎来源的间充质干细胞中的一种或多种。
进一步地,所述间充质干细胞为骨髓间充质干细胞。
进一步地,所述骨质疏松症为原发性骨质疏松症或继发性骨质疏松症。
进一步地,所述外泌体制剂用于增加骨密度。
进一步地,所述外泌体制剂为注射剂、胶囊剂、片剂、粉末剂、软膏剂或喷雾剂等的形式。
进一步地,所述药学上可接受的载体包括磷酸盐缓冲液和/或生理盐水。
另一方面,本发明提供间充质干细胞外泌体制剂在制备用于治疗或预防骨质疏松症的药物、保健品或营养补充剂中的应用。
又一方面,本发明提供间充质干细胞外泌体制剂在制备用于增加骨密度的药物、保健品或营养补充剂中的应用。
本发明的有益效果包括:本发明的间充质干细胞外泌体制剂制备简单、材料来源广泛、且相较于微米级的细胞,外泌体囊泡的提取方法更为简单灵活且容易保存。通过施用间充质干细胞外泌体,显著地提高了骨密度,从而实现预防和治疗骨质疏松症的目的。另外,本发明的外泌体制剂不具有副作用或副作用小、且不会引起患者不适反应,因此能够有效地长期用于预防或治疗骨质疏松症。
附图说明
图1示出骨髓间充质干细胞的生长形态光镜图;
图2示出骨髓间充质干细胞外泌体的透射电镜图,其示出微囊泡样结构;
图3示出Western blot鉴定骨髓间充质干细胞外泌体的表面标志物的结果;
图4示出利用NaNoSight LM10显微镜系统鉴定的外泌体直径大小的分布;
图5A-5C示出不同浓度的外泌体处理对骨质疏松症治疗的效果图;和
图6A-6C示出外泌体和乙酰水杨酸组合处理对骨质疏松症治疗的效果图。
具体实施方式
下面结合实施例,进一步阐述本发明。应理解,本发明的技术方案不限于以下所列举的具体实施方式,还包括各种具体实施方式的组合。
下述实施例中所使用的实验方法如无特殊说明,均为常规方法。
下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。
实施例1:骨髓间充质干细胞外泌体的制备
1、骨髓间充质干细胞的分离和培养
1)取6-8周龄小鼠股骨和胫骨,将表面附着的肌肉及筋膜等清除干净,并且用PBS冲洗;
2)将股骨和胫骨两端断开,用10ml注射器抽取含2%热灭活胎牛血清的PBS从干骺端反复冲洗骨髓腔,然后100μm细胞筛过滤,获得骨髓细胞;
3)将所得骨髓细胞离心(1500r/min)5min,然后用培养液重悬离心后所得的骨髓细胞,再将其接种于直径100mm的培养皿中,并置于37℃、5%CO2的培养箱中培养,24小时后更换培养液;
4)当细胞达到70-80%融合时进行传代培养,用于间充质干细胞鉴定或后续的外泌体分离,其中骨髓间充质干细胞细胞生长状态如图1所示。
其中,所述培养液包括:含20%胎牛血清(FBS)、2mM谷氨酰胺、55μM巯基乙醇、100U/ml青霉素和100μg/ml链霉素的α-MEM。
2、骨髓间充质干细胞的鉴定
(1)骨髓间充质干细胞表面标记分子的测定
利用荧光标记的抗体CD105、CD90、SCA1、CD34和CD45对上述获得的骨髓间充质干细胞进行染色30min,在用2%PFA固定后,利用流式细胞仪(BD Accuri C6)检测干细胞表面标记分子表达,其中骨髓间充质干细胞高表达CD105(93.39%)、CD90(72.44%)和SCA1(44.57%),但几乎不表达造血干细胞表面标记分子CD34(0.05%)和CD45(0.01%)。
(2)细胞增殖的测定
将上述间充质干细接种于4孔细胞玻片(Nunc,1×104/孔)上,在含5%CO2、温度为37℃的培养箱中培养12小时。通过溴脱氧尿苷方法检测细胞增殖率。
在细胞培养物中加入溴脱氧尿苷(10μM),继续培养12小时。移除培养液,PBS冲洗三次细胞培养物,然后以70%乙醇固定30分钟。移除固定液,蒸馏水冲洗两次,每次2分钟。加入变性液(1.5M HCl,100μl)孵育30分钟,移除变性液。PBS冲洗三次,每次2分钟。加入封闭液(5%BSA溶液,100μl)孵育10分钟。移除封闭液,加入生物素耦联的小鼠抗溴脱氧尿苷抗体孵育60分钟。移除抗体,PBS冲洗三次,每次2分钟。加入链霉亲和素耦联的过氧化酶孵育10分钟。移除过氧化物酶,加入DAB显色剂(100μl)显示阳性染色,苏木素(100μl)复染细胞核,显示间充质干细胞快速增殖。
(3)成骨分化检测
在成骨培养液(含1.8mM磷酸二氢钾,10nM地塞米松)中诱导间充质干细胞进行成骨分化。经过28天的成骨诱导,用茜素红对钙结节进行染色,用免疫印迹分析成骨标记物碱性磷酸酶(ALP和RUNX2)的表达。
(4)成脂分化检测
在成脂培养液(含0.5mM异丁甲基黄嘌呤、60μM吲哚米辛、0.50μM氢化可的松和100μg/ml胰岛素)中诱导间充质干细胞进行成脂分化。经过14天的成脂诱导,用油红O对脂滴进行染色,用半定量PCR分析成脂标记物LPL和PPAR的表达。
3、外泌体的分离和鉴定
(1)外泌体的分离
A.将骨髓间充质干细胞在无外泌体的培养基中扩增3天,收集培养基;
B.将收集到的细胞培养基在15℃以2000rpm离心30min去除培养基中的细胞碎片;
C.弃去沉淀,收集上清液;
D.对上清液进行系列超速离心以获得外泌体,具体地:1)300g离心10分钟,收集上清液;2)2,000g离心10分钟,收集上清液;3)10,000g离心10分钟,收集上清液;4)100,000g离心70分钟,收集沉淀;5)将沉淀用PBS重悬,100,000g离心70分钟,收集的沉淀部分即为分离的外泌体。
(2)外泌体的鉴定
通过透射电镜(FEI Tecnai Spirit 120kv)观察所分离的骨髓间充质干细胞,发现细胞内存在大量的微囊泡样的外泌体(见图2)。
用常规方法提取外泌体蛋白,通过Western blot检测外泌体中表面标志物CD9和CD81的表达(见图3)。
外泌体直径的测定:
1)由培养基分离得到的细胞外囊泡重悬于PBS中;
2)经不同比例的稀释,利用NanoSight LM10显微镜系统,记录细胞外囊泡的形态,并进行直径大小的测量。如图4所示,骨髓间充质干细胞分离得到的外泌体囊泡的尺寸集中在40-250nm范围内,特别地在200nm附近。
实施例2:骨髓间充质干细胞外泌体对骨质疏松症的作用
(1)小鼠模型:取6-8周龄的雌性小鼠C3H小鼠,体重20g左右,作为实验小鼠。通过卵巢去势法,构建小鼠骨质疏松症状模型。同时建立假手术(Sham)组作为对照,该组仅去除少许卵巢周围脂肪。实验小鼠均从北京维通利华购买。
(2)分组试验:建立小鼠骨质疏松症模型4周后,将小鼠随机分为四组:OVX组(仅0.9%NaCl处理)、Exo-100μg组(骨髓间充质干细胞外泌体100μg溶于200μl 0.9%NaCl中)、Exo-200μg组(骨髓间充质干细胞外泌200μg溶于200μl0.9%NaCl中)和Exo-500μg组(骨髓间充质干细胞外泌体500μg溶于200μl0.9%NaCl中),每组6只小鼠。通过尾静脉分别向三组小鼠注射100μg、200μg和500μg骨髓间充质干细胞外泌体(200μl)进行治疗,OVX组用同样的方法仅注射200μl 0.9%NaCl溶液,注射四周后,观察结果。
结果表明:如图5A-5C所示,骨髓间充质干细胞外泌体治疗小鼠的骨密度(BMD)和骨体积分数(BV/TV)相较于OVX组均呈现增加趋势。其中Exo-200μg治疗组对小鼠骨密度和骨体积分数的提高效果显著优于Exo-100μg和Exo-500μg治疗组(P<0.05)。这表明施200μg外泌体的治疗效果要显著地优于施用100μg和500μg外泌体的治疗效果。
实施例3:骨髓间充质干细胞外泌体和乙酰水杨酸组合对骨质疏松症的作用
在实施例2的结果基础上,进一步探讨了外泌体与乙酰水杨酸联合治疗对骨质疏松症的影响。
根据实施例2中的操作步骤,对小鼠注射骨髓间充质干细胞和乙酰水杨酸(acetosalicylic acid,ASA)以进行联合治疗。其中,具体治疗分组如下:OVX组(仅200μl的0.9%NaCl处理)、ASA组(仅200μl的浓度为200μg/ml的ASA)、Exo组(200μl的浓度为1000μg/ml的骨髓间充质干细胞外泌体)和ASA+Exo组(200ul的1000μg/ml骨髓间充质干细胞外泌体+浓度为200μg/ml的ASA),每组6只小鼠。通过尾静脉分别对小鼠进行如上处理,注射四周后,观察结果。结果表明,外泌体和乙酰水杨酸联合治疗组的骨密度图像以及骨密度和骨体积分数与假手术组(无骨质疏松症)几乎相同,且显著地优于OVX组、单纯地外泌体组或者单纯地乙酰水杨酸组治疗的效果。这充分说明外泌体和乙酰水杨酸组合对于骨质疏松症的治疗呈现出明显的协同作用,其显著提高外泌体对骨质疏松症治疗的效果(如图6A-6C所示)。
以上所述仅仅是本发明的优选实施方式。应当指出的是,在不脱离本发明的精神和实质的情况下,本领域技术人员可对本发明的细节进行各种修改、变更或替换。这些修改、变更或替换也应理解为包括在本发明要求保护的范围之内。
Claims (8)
1.一种用于治疗骨质疏松症的间充质干细胞外泌体制剂,由骨髓间充质干细胞来源的外泌体、药学上可接受的载体和乙酰水杨酸组成;
所述外泌体和所述乙酰水杨酸的质量比为1:1至3:1。
2.根据权利要求1所述的制剂,其中,相对于所述外泌体制剂的总体积,所述外泌体制剂中的外泌体含量为500μg/ml至2500μg/ml。
3.根据权利要求1-2中任一项所述的制剂,其中,所述制剂包含100μg至500μg的外泌体。
4.根据权利要求1所述的制剂,其中,所述外泌体的直径为40nm至250nm。
5.根据权利要求1所述的制剂,其中,所述骨质疏松症为原发性骨质疏松症或继发性骨质疏松症。
6.根据权利要求1所述的制剂,其中,所述外泌体制剂为注射剂、胶囊剂、片剂、粉末剂、软膏剂或喷雾剂的形式。
7.根据权利要求1所述的制剂,其中,所述药学上可接受的载体包括磷酸盐缓冲液和/或生理盐水。
8.根据权利要求1-7中任一项所述的制剂在制备用于治疗骨质疏松症的药物中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910867109.8A CN110403959B (zh) | 2019-09-12 | 2019-09-12 | 间充质干细胞外泌体制剂及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910867109.8A CN110403959B (zh) | 2019-09-12 | 2019-09-12 | 间充质干细胞外泌体制剂及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN110403959A CN110403959A (zh) | 2019-11-05 |
CN110403959B true CN110403959B (zh) | 2021-05-11 |
Family
ID=68370303
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910867109.8A Active CN110403959B (zh) | 2019-09-12 | 2019-09-12 | 间充质干细胞外泌体制剂及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN110403959B (zh) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114073714A (zh) * | 2020-08-11 | 2022-02-22 | 北京大学口腔医学院 | 干细胞外泌体制剂及其应用 |
CN112107599B (zh) * | 2020-08-14 | 2022-06-21 | 深圳市弘际生物科技有限责任公司 | 干细胞外泌体在制备用于治疗膀胱过度活动综合症的药物中的应用 |
CN112022876A (zh) * | 2020-09-01 | 2020-12-04 | 华中科技大学同济医学院附属协和医院 | 骨髓间充质干细胞外泌物在椎间盘退行性疾病中的应用 |
CN115011554B (zh) * | 2022-06-14 | 2023-04-18 | 上海依赛洛森生物医药有限公司 | 骨髓间充质干细胞的外泌体、体外培养方法以及应用 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108478600B (zh) * | 2018-05-04 | 2024-01-30 | 中南大学湘雅医院 | 间充质干细胞外泌体在制备防治骨质疏松症药物中的应用 |
-
2019
- 2019-09-12 CN CN201910867109.8A patent/CN110403959B/zh active Active
Also Published As
Publication number | Publication date |
---|---|
CN110403959A (zh) | 2019-11-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN110403959B (zh) | 间充质干细胞外泌体制剂及其应用 | |
Hoang et al. | Differential wound healing capacity of mesenchymal stem cell-derived exosomes originated from bone marrow, adipose tissue and umbilical cord under serum-and xeno-free condition | |
Di Taranto et al. | Qualitative and quantitative differences of adipose-derived stromal cells from superficial and deep subcutaneous lipoaspirates: a matter of fat | |
Guo et al. | Exosomes from human synovial-derived mesenchymal stem cells prevent glucocorticoid-induced osteonecrosis of the femoral head in the rat | |
CN105861430A (zh) | 一种外泌体、外泌体的制备方法及其在制备治疗脓毒症药物或者制剂中的应用 | |
CN109432126A (zh) | 与间充质干细胞外来体相关的方法和组合物 | |
US11447748B2 (en) | Encapsulated adipose-derived stem cells, methods for preparation and theraputic use | |
CN113136362B (zh) | 一种囊泡及其应用 | |
JP5431146B2 (ja) | 免疫学的寛容および調節性前駆細胞 | |
TW200902718A (en) | Procurement, isolation, and cryopreservation of endometrial/menstrual cells | |
WO2021147923A1 (zh) | 一种囊泡及其应用 | |
KR20180054523A (ko) | 외래 미토콘드리아를 포함하는 자연살해세포 및 이를 포함하는 약학적 조성물 | |
CN103301154B (zh) | 脐带间充质干细胞在制备治疗红斑狼疮的制剂中的用途 | |
CN113712995A (zh) | 神经干细胞联合脐带间充质干细胞在脊髓损伤中的应用 | |
JP6722598B2 (ja) | 関節リウマチ治療のための間葉系間質細胞 | |
Chen et al. | Small extracellular vesicles from human adipose-derived mesenchymal stromal cells: a potential promoter of fat graft survival | |
CN115478048A (zh) | 培养脂肪间充质干细胞制备外泌体 | |
Hoogduijn et al. | Donor-derived mesenchymal stem cells remain present and functional in the transplanted human heart | |
CN103865873B (zh) | 亚全能干细胞分泌的外来体及其应用 | |
CN108486039B (zh) | 小分子诱导人脂肪干细胞分化为睾丸间质细胞的方法 | |
Kitala et al. | Possibilities and Limitations in Current Translational Stem Cell Research | |
JP7003283B2 (ja) | 末梢血から間葉系幹細胞集団を製造する方法及びその使用 | |
CN113215100B (zh) | 小分子化合物mln4924在促进外泌体分泌中的应用 | |
Samoylovich et al. | The influence of mesenchymal stromal cells on B-cell line growth and immunoglobulin synthesis | |
Jin et al. | Differentiation of Human Umbilical Cord Derived Mesenchymal Stem Cells Into Neural Stem Cells Induced by hPRDX5 |
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 |