人脐带间充质干细胞的分离培养与鉴定简

目的：建立并优化人脐带间充质干细胞分离纯化方法，并对其表面标志与多向分化潜能进行鉴定。方法：收集健康足月产胎儿脐带组织，采用组织块贴壁法进行原代培养，流式细胞仪对其表面标志进行检测，通过向成骨成脂分化对其多向分化潜能进行鉴定，RT-PCR对其干细胞特性基因Oct4、Nanog、Sox2、Nestin进行检测。结果：采用组织块贴壁法可在2周左右获得大量间充质干细胞，培养的细胞经流式细胞仪检测，高表达CD29、CD44、CD105、CD106，低表达CD34、CD45；经成骨成脂诱导2周后可分化为成骨细胞和成脂细胞，RT-PCR检测发现原代细胞表达Oct4、Nanog、Sox2、Nestin基因。结论：人脐带间充质干细胞可在体外扩增培养，具有多向分化潜能，可作为组织工程种子细胞来源。

Isolation and Identification of Human Umbilical Cord Mesenchymal Stem Cells

Objective： To establish and optimize the method of isolation of human mesenchymal stem cells （MSC） and identify their surface markers and mulitidifferentiation ability. Methods： The MSC were isolated from neonate umbilical cord, cultured by tissue explants adherent method. The surface markers were identified by flow cytometry, multi-differentiation capacity was identified by osteogenic and adipogenic differentiation, stem cell mark er gene Oct4, Nanog, Sox2 and Nestin were detected by RT-PCR. Results： A substantial number of MSC can be harvested using the tissue explants adherent method at about two weeks, CD29, CD44, CD105 and CD106 highly expressed, while CD34, CD45 weakly expressed on the ceils surface through the detection of flow cytometry, after induction for 2 weeks. These cells can differentiated into adipogenic and osteogenic ceils. RT-PCR results showed that these cells expressed Oct4, Nanog, Sox2 and Nestin. Conclusion： The human umbilical cord MSC can be cul tured and proliferated in vitro, these cells can be used as a new source of tissue engineering due to their capacity of muhidifferentiation.