【摘 要】
:
目的:分离人骨髓间充质干细胞(hMSCs),建立克隆化扩增的细胞系并初步鉴定其分化特性和细胞的分子标记.方法:利用间充质干细胞贴壁生长的特性,显微镜下挑取原代单个成纤维样生
【机 构】
:
第二军医大学基础医学部细胞生物学教研室,The Department of Molecular and Cellular Biology
论文部分内容阅读
目的:分离人骨髓间充质干细胞(hMSCs),建立克隆化扩增的细胞系并初步鉴定其分化特性和细胞的分子标记.方法:利用间充质干细胞贴壁生长的特性,显微镜下挑取原代单个成纤维样生长单位中的细胞,逐步扩大培养,最终得到克隆化扩增的hMSCs.选择有利于其生长的血清,低密度培养,传70%~80%生长汇合时传代保种.RT-PCR检测其Oct-4、SDF1、CD49a、CK19、c-met基因的表达;流式细胞术检测细胞表面抗原CD34、CD45、CD14、CD44、CD29、CD90、HLA-1和HLADR;体外向骨、软骨、脂肪细胞方向诱导分化鉴定其分化潜能.结果:建立了人骨髓间充质干细胞系并在体外实现了克隆化扩增,该细胞系在体外连续培养达60个细胞倍增时间仍保持多向分化的潜能.细胞不表达CD34、CD45、CD14、HLA-DR,但表达Oct-4、SDF-1、CD49a、CK19、c-met、CD44、CD29、CD90、HLA-1.体外能诱导出骨、软骨、脂肪细胞.结论:得到了一个可稳定传代的克隆化扩增的人骨髓间充质干细胞系,该细胞系在体外可以分化为骨、软骨、脂肪细胞,并表达Oct-4、CK19和c-met.
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