摘要
目的从胎儿骨髓中分离培养纯化骨髓间充质干细胞(MSC),并进行细胞表面标志的检测,探讨MSC的培养方法。方法采用全骨髓培养、贴壁筛选法分离培养MSC,采用差速贴壁培养方法纯化细胞,采用流式细胞仪检测细胞表面标志,并观察不同方法处理的培养界面对细胞贴壁和生长的影响。结果通过分离纯化培养,光镜下细胞形态以长梭形、不规则多角形为主;IV型胶原包被和Co60照射(照射量6GY)处理的培养瓶更利于MSC的贴壁、生长;流式细胞仪检测结果显示CD44(++)、Stro-1(+)、CD14(-)、CD34(-)、CD45(-)、CD11b(-)。结论采用贴壁筛选法和差速贴壁法结合可获得较高纯度的MSC,并且细胞生长良好,在体外具有较强的扩增能力。
Objective To develop the methodology of the culture of mesenchymal stem cell （MSC） derived from bone marrow of human embryo. Methods Adherent cells were selected as MSCs after the whole marrow was cultured, and the method of different adherence speed was applied to isolate and purify MSC. The expression of surfaee molecules was examined by flow cytometry. The effect of various treatment af culture flask bottom on adherence and cell growth was observed. Results The isolated MSCs showed long spindle and multiangular shapes under contrast phase microscopy. Culture flask covered by collagen type Ⅳ or irradiated by Co60 （6GY） was better for adherence and cell growth. Flow cytometric analysis demonstrated positive expression of CD44（＋＋）,Stro- 1 （＋）, CD 14（-）, CD34（-）, CD45（-）and CD 11 b（-）on the cell surfaee. Conclusion MSC with high purity can be obtained by adherent screening method combined with the method of different adherence speed in vitro.
出处
《浙江医学》
CAS
2007年第12期1265-1267,共3页
Zhejiang Medical Journal
基金
温州市科委资助课题(S2002A130)
浙江省自然科学基金课题(YZ6679)
