人脐血间充质干细胞分离培养方法的优化  

OPTIMIZATION OF THE ISOLATION AND CULTIVATION METHOD OF HUMAN UMBILICAL CORD BLOOD MESENCHYMAL STEM CELLS

作　　者：张炳强[1] 于丽[1] 石娟[1] 管英俊[1] 马丽[2] 张雪莉[1] 

机构地区：[1]潍坊医学院组织学与胚胎学教研室,潍坊261042 [2]潍坊市妇幼保健院,潍坊261011

出　　处：《中国组织化学与细胞化学杂志》2009年第2期123-127,共5页Chinese Journal of Histochemistry and Cytochemistry

基　　金：山东省教育厅基金资助项目(J07YD01);潍坊医学院博士启动基金

摘　　要：目的探讨人脐血间充质干细胞(MSCs)体外培养纯化的最佳方法,为脐血MSCs在临床的广泛应用奠定基础。方法无菌条件下采集足月分娩和早产儿脐血,密度梯度离心法分离脐血单个核细胞,比较胎龄、不同培养基、接种密度、首次换液时间对脐血MSCs原代培养过程的影响,通过免疫荧光方法检测表面标记物的表达情况,观察脐血MSCs的生物学特性。结果足月分娩脐血,采用MesencultTM培养基,以5×106/cm2的密度接种,首次换液时间为7d时,脐血MSCs原代培养成功率较高。相同培养条件下,早产儿脐血培养成功率高于足月分娩脐血。人脐血MSCs强表达CD29、CD44和CD90,不表达造血干细胞表面标志CD34。结论优化筛选到一种合适的人脐血MSCs培养纯化条件。Objective To explore the optimal condition of isolation and cultivation of mesenchymal stem cells （MSCs） derived from human umbilical cord blood. Methods Umbilical cord blood was collected from full-term deliveries and preterm infants. The umbilical cord blood mononuclear cells were isolated by density gradient centrifugation with a lymphocyte separation medium. The influence of different gestational ages, culture media, planting densities and the time points of the first medium change on the growth of MSCs were analyzed. The biological characteristics of MSCs were observed by detecting the cell surface markers with immunofluorescence. Result When the other conditions were the same, MesencultTM medium was the best medium, 5 × 10^6/cm^2 was the best planting density, the best time for the first medium change was on the seventh day. The achievement ratio of umbilical cord blood from preterm infants was higher than that of full-term deliveries when the other conditions were the same. MSCs derived from human umbilical cord blood expressed CD29, CD44 and CD90, but not antigens of hematopoietic CD34. Conclusion We have found the optimized method of culture of umbilical cord blood mesenchymal stem cells.

关 键 词：脐血 间充质干细胞 细胞培养 优化 

分 类 号：R331.1[医药卫生—人体生理学]