密度梯度法与羟乙基淀粉法培养脐血间充质干细胞比较  

Comparison of two separation methods of culturing human umbilical cord blood mesenchymal stem cells

作　　者：郎长会 彭龙英[1] 束晓梅[1] 

机构地区：[1]遵义医学院附属医院儿科,贵州遵义563099

出　　处：《遵义医学院学报》2013年第5期477-480,共4页Journal of Zunyi Medical University

基　　金：国家自然科学基金资助项目(NO:31260286)

摘　　要：目的比较密度梯度离心法与羟乙基淀粉法体外分离培养脐血间充质细胞(hUCB-MSCs)的成功率,为今后细胞移植和基因治疗提供重要的细胞来源。方法取44份足月儿顺产或剖宫产脐带血,分为两组,每组22份,分别用密度梯度离心法结合贴壁法与羟乙基淀粉法分离脐血中单个核细胞,专用培养基培养,观察两种方法培养所得细胞的形态变化并绘制生长曲线,用流式细胞术鉴定细胞表面标志物CD14,CD29,CD44,CD45,CD105的表达。结果羟乙基淀粉法培养hUCB-MSCs的成功率(36.36%)高于密度梯度离心法结合贴壁法的培养成功率(9.1%)(P<0.05)。两种方法所得细胞在形态、生长曲线和表面标志物CD14,CD29,CD44,CD45,CD105的表达无明显差异(P>0.05)。结论羟乙基淀粉法可以提高hUCB-MSCs培养成功率,为hUCB-MSCs在临床中应用奠定基础。Objective To explore an optimized isolation method of human umbilical cord blood mesen- chymal stem cells （hUCB -MSCs）. Methods Umbilical cord blood cells were collected from full -term normal delivery or cesarean section delivery infants under aseptic condition. 44 samples of umbilical cord blood were collected, and divided into two groups. The density gradient centrifugation or hydroxy- ethyl starch sedimentation was used to isolate hUCB -MSCs with adherent culture, and compared. Mononuclear cells （MNCs） were harvested from human umbilical cord blood. The morphology of MNC and their growth characteristics were observed. The surface antigen phenotype was detected by flow cy- tometry. Results The success rate of the hydroxyethyl starch sedimentation method was higher than that of density gradient centrifugation （P 〈 0. 05 ）. The morphological changes and the surface markers CD14, CD29, CD44, CD45 and CD105 were not obviously different （ P 〉 0.05）. Conclusion The hydroxyethyl starch sedimentation culture could improve the success rate of hUCB -MSCs.

关 键 词：密度梯度离心 贴壁筛选法 羟乙基淀粉 脐血 间充质干细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]