人脐血间充质干细胞体分离、培养及向脂肪细胞分化的实验研究

Study on the isolation cultured of human cord blood mesenchymal stem cells and its induction differentiation into adipocytes

目的:探讨脐血间充质干细胞(UCB-MSCs)的分离培养及向脂肪细胞的分化潜能。方法:从足月儿脐血中获得间充质干细胞,体外培养传代,流式细胞检测细胞表面及抗原细胞周期;予10-6M地塞米松、100μg/ml1-甲基-3-异丁基-黄嘌呤(IBMX)、50μg/ml抗坏血酸的IMDM培养基,对MSCs进行诱导向脂肪细胞分化,油红-O染色染色鉴定。结果:成功建立了脐血间充质干细胞分离及培养扩增的方法;流式细胞仪检测结果显示,贴壁细胞均表达CD105、CD29和CD44,不表达造血细胞表型CD34、CD45和内皮细胞表型CD31;经脂肪培养液诱导后在细胞胞浆里可见颗粒状红色脂滴形成。结论:脐血间充质干细胞能进行体外分离培养,并能诱导分化为脂肪细胞。

Objective：To establish a method to separate mesenchymal stem cells from human umbilical cord blood（UC-MSCs）,culture UC-MSCs in vitro,and study its biological characteristics.Methods：MSCs were seperated from human umbilical cord blood and expanding cultured in vitro,Surface antigens were detected with flow cytometry;MSCs were induced to differentiate into adipocytes in induction medium containing dexmethasone,IBMX and ascorbic acid.Then differentiated cells were identificated oil red O stain.Results：The method of culture and expansion of UCB-M SCs in vitro were established.Adhesive cells were all positive for MSC-related antigens,such as CD29,CD105 and CD44,but negative for CD34,CD45 and CD31.UCB-MSCs differentiated into adipocytes and lipid droplets were scarlet red by oil red O stain on the 14th day in adipose medium.Conclusion：The cultured cells,with the biological characteristics of MSCs,can be amplified easily,passaged and differentiate into adipocyte in vitro.