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人骨髓间充质干细胞体外分离培养方法的改进

Modified Isolation and Cultivation of Human Mesenchymal Stem Cells in Vitro
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摘要 目的:对人骨髓间充质干细胞(Mesenchymalstemcells,MSCs)的体外分离纯化、培养扩增方法的改 进,为MSCs的进一步诱导分化和应用奠定基础。方法:抽取人骨髓细胞,采用密度梯度离心结合贴壁培养法 进行分离纯化,并传代扩增,测定生长曲线和贴壁率,形态学观察,流式细胞仪检测其表面抗原表达。结果:密 度梯度离心结合贴壁培养法能有效分离纯化人骨髓MSCs,MSCs在含胎牛血清的L DMEM培养液中生长性状 相对稳定,细胞基本呈成纤维细胞样生长,1、3、5代细胞生长曲线、贴壁率基本相同,贴壁存活率高。表达 CD29、CD44、CD105、CD166。结论:密度梯度离心结合贴壁培养法能有效分离纯化人骨髓MSCs,在含胎牛血清的 L DMEM培养液中,细胞稳定扩增。 Objective:To modify the isolation and cultivation of mesenchymal stem cells(MSCs) from human bone marrow for offering an experimental foundation for their further differentiation and actual application. Methods: Isolated and purified the human marrow MSCs by density gradient centrifugation and culture plastic-adhering. After in vitro subculture and expansion, observed the morphology of the cells, drew the growth curve, measured the adhesive rate, analyzed MSCs by flow cytometry. Results: The living characteristic was stable in L-DMEM medium containing newborn bovine serum. The growth curves and adhesive rates of passage 1, 3, and 5 were quite similar. MSCs exhibited a typical fibroblastlike morphology and expressed CD 29, CD 44, CD 105, CD 166. Conclusion: The marrow MSCs could be effectively isolated and purified by density gradient centrifugation and culture plastic-adhering and expanded satisfactorily in L-DMEM with newborn bovine serum.
出处 《广州医学院学报》 2004年第4期54-57,共4页 Academic Journal of Guangzhou Medical College
基金 广东省自然科学基金(04009683) 广州市教育局资助项目(2004-1003)
关键词 骨髓间充质干细胞 分离培养 体外 mesenchymal stem cells isolation and cultivation human in vitro
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  • 1鄂征.组织与细胞培养技术(第2版)[M].北京:北京人民出版社,1995.9-20.
  • 2Bruder SP, Fink DJ, Caplan AI. Mesenchymal stem cells in bone development, bone repair, and skeletal regeneration therapy [J]. J Cell Biochem,1994;56(3):283-294.
  • 3Aubin JE. Bone stem cells [J]. J Cell Biochem,1998;30/31(Suppl):73-82.
  • 4Yamaguchi A, Ishizuya T, Kinto N. Effects of BMP-2, BMP-4, and BMP-6 on osteoblastic differentiation of bone marrow-derived stromal cell lines, ST2 and MC3T3-G2/PA6 [J]. Biochem Biophys Res Commun, 1996;220(2):366-371.
  • 5Locklin RM, Williamson MC, Beresford JN, Owen ME. In vitro effects of growth factors and dexamethasone on rat marrow stromal cells [J]. Clin Orthop, 1995;313:27-35.
  • 6Beresford JN, Graves SE, Smoothy CA. Formation of mineralized nodules by bone derived cells in vitro: A model of bone formation [J]? Am J Med Genet, 1993;45(2):163-178.
  • 7Hasegawa Y, Ohgushi H, Ishimura M, Habata T, Tamai S, Tomita N, Ikada Y. Marrow cells culture on poly-L-lactic acid fabrics [J]. Clin Orthop, 1999; 358:235-243.
  • 8Verfaillie C M,J Exp Med,1994年,179卷,643页
  • 9Huang S,Nature,1993年,360卷,745页
  • 10魏宽海.组织工程化骨组织中成骨细胞来源的选择[J].国外医学(创伤与外科基本问题分册),1998,19(3):155-158. 被引量:23

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