足月小样儿及正常胎儿脐血间充质干细胞的分离培养及形态学观察

The isolation and morphological observation of umbilical cord blood mesenchymal stem cells from preterm infant

目的建立一种简单实用的脐血间充质干细胞(MSCs)的分离培养方法,观察其生物学特性,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小样儿和足月正常胎儿分娩脐血,密度梯度离心法分离脐血单个核细胞,采用MesencultTM培养基培养,观察脐血MSCs的生物学特性,免疫荧光方法检测其表面标记物的表达情况。结果采用MesencultTM培养基,正常儿脐血MSCs培养成功率为66.67%,相同培养条件下,足月小样儿脐血培养成功率低于正常体重胎儿的脐血。人脐血MSCs强表达CD29、CD44和CD90,不表达造血干细胞表面标志CD34。结论我们建立了最佳的脐血MSCs的分离培养方法。

Objective：To explore a simple and pragmatic way of isolation and cultivation of mesenchymal stem cells（MSCs） derived from preterm infant umbilical cord blood.Methods：Umbilical cord blood were collected from preterm infant and full term deliveries.The umbilical cord blood mononuclear cells were isolated by density gradient centrifugation with lymphocyte separation medium.Using MesencultTM culture medium,the biological characteristics of MSCs derived from human umbilical cord blood were observed by detecting the cell surface marker by immunofluorescence method.Results：MesencultTM medium was the best medium,the achievement ratio of umbilical cord blood from preterm infant was 100%,higher than 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 from preterm infant.