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华西医学  2015 

人脐带间充质干细胞的体外获取、鉴定及标记

DOI: 10.7507/1002-0179.20150586, PP. 2067-2070

Keywords: 间充质干细胞,脐带,,分离,培养,分化

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Abstract:

目的?从人脐带中获取间充质干细胞(MSC)并进行标记,为进一步移植治疗奠定基础。方法?运用组织块贴壁法从人脐带中分离MSC,通过传代培养后,进行形态学观察;采用流式细胞仪对人脐带间充质干细胞(hUC-MSC)的细胞表型进行鉴定;通过特异性染色鉴定体外成脂、成骨诱导后的分化能力;运用免疫荧光证实BrdU标记细胞是否成功。结果?运用组织块贴壁法可成功从人脐带中分离出MSC,培养1周左右可见细胞从组织块边缘爬出,向外增殖生长,形成集落;流式细胞仪检测P5代细胞,高表达CD73、CD90及CD105,不表达或低表达CD14、CD34、CD45、CD79a及人类白细胞抗原-DR;成脂诱导2周,油红O染色为阳性,成骨诱导3周,茜素红染色出现红色沉淀物;BrdU标记细胞经免疫荧光检测后,可见红色荧光。结论?运用组织块贴壁法从人脐带中分离的细胞,具备MSC的特点并可成功被标记,可用于细胞移植领域的研究。

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