脐带源性和脐血源性间充质干细胞分离培养方法的比较

目的建立分离培养人脐带间充质干细胞（UCMSCs）和脐带血间充质干细胞（UCBMSCs）的方法，并比较其效果。方法无菌条件下采集健康足月新生儿脐带及脐血各30份，分别通过原代贴壁培养法、酶消化法培养UCMSCs，及淋巴细胞分离液法、羟乙基淀粉沉降与淋巴细胞分离两步分离法获得脐血中单个核细胞培养UCBMSCs。应用含10％胎牛血清的DMEM／F12培养基，比较UCMSCs和UCBMSCs分离培养的效果与生长形态。流式细胞术检测其表面标志及诱导分化鉴定其分化能力。结果UCMSCs原代贴壁培养法8d左右可见成纤维样细胞从组织块边缘爬出且成簇生长，酶消化法培养UCMSCs5d左右均匀生长；UCBMSCs分离培养用羟乙基淀粉沉降与淋巴细胞分离两步法得到的细胞数量较淋巴细胞分离法明显增多。两种来源MSCs培养方法相比较，UCMSCs原代培养的时间短，培养成功率明显增高。流式细胞仪检测两种来源的MSCs具有MSCs表面标志特征。定向诱导分化结果表明MSCs具有被诱导为成骨细胞、脂肪细胞的分化能力。结论人脐带来源间充质干细胞原代培养周期短，培养效率更高；脐血间充质干细胞的分离方法中羟乙基淀粉沉降与淋巴细胞分离两步法效率更高。

Comparison of Isolation and Culture Methods between Umbilical Cord Mesenchymal Stem Cells and Umbilical Cord Blood Mesenchymal Stem Cells

Objective To establish methods for isolation and culture of umbilical cord mesenchymal stem cells(UC MSCs) and umbilical cord blood mesenchymal stem cells(UCB MSCs) in vitro,and compare the effects.Methods Umbilical cord blood(n=30) and Umbilical cord(n=30) were collected from healthy full-term deliveries.Human UC MSCs were isolated and cultured using adherence technique and enzyme digestion,and mononuclear cells were isolated from umbilical cord blood with the lymphocyte isolation method and hydroxyethyl s...