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何晓婷,李怀芳,王建军,等.人脐血间充质干细胞的改良分离培养[J].同济大学学报（医学版）,2012,33(2):33-37. [点击复制]
HE Xiao-ting,LI Huai-fang,WANG Jian-jun,et al.Improved method for isolation and culture of mesenchymal stem cells from human umbilical cord blood[J].同济大学学报（医学版）,2012,33(2):33-37. [点击复制]

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人脐血间充质干细胞的改良分离培养
何晓婷¹,李怀芳¹,王建军¹,孙向华²
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(1. 同济大学附属同济医院妇产科,上海,200065;2. 同济大学附属同济医院中心实验室,上海,200065)

摘要:

目的探讨人间脐血来源的充质干细胞在体外分离培养与扩增的方法。方法用重力离心-密度梯度离心法结合贴壁筛选法分离细胞,用含15%FBS的DMEM低糖培养基培养并传代,流式细胞仪检测P3代间充质干细胞的表面标志。结果脐血来源的单个核细胞24 h即开始贴壁,主要表现为破骨样细胞和间充质样细胞。经自然纯化法传代后,可得到形态单一的长梭形间充质干细胞。流式细胞仪检测结果显示该细胞表达CD44。结论脐血来源的MSCs可以在体外分离培养和扩增,细胞表面标记与骨髓来源间充质干细胞一致。

关键词: 人脐血间充质干细胞分离培养

DOI：10.3969/j.issn1008-0392.2012.02.007

基金项目:上海市科学技术委员会科研计划项B(09411%3700)

Improved method for isolation and culture of mesenchymal stem cells from human umbilical cord blood

HE Xiao-ting¹,LI Huai-fang¹,WANG Jian-jun¹,SUN Xiang-hua²

(1.Dept.of Gynecology and Obstetrics,Tongji Hospital,Tongji University,Shanghai 200065,China;2.Central Laboratory,Tongji Hospital,Tongji University,Shanghai 200065,China)

Abstract:

Objective To improve the method for isolation and culture of human umbilical cord blood-derived mesenchymal stem cells(hUCB-MSCs) in vitro.Methods The cord blood mononuclear cells were isolated by density gradient centrifugation and directly adherence growth method.After cultured with DMEM containing low glucose and 15% FBS medium for 3 passages,surface antigen expression of MSCs was detected by flow cytometry.Results The cord blood mononuclear cells began to adherent to flasks in 24h,mainly displayed osteoblast-like cells and mesenchymal-like cells.By natural purification for 3 passages,long spindle shaped-MSCs were harvested.Test results from flow cytometry showed that the cells expressed CD44 strongly.Conclusion With the improved method hUCB-MSCs can be isolated,cultured and expanded in vitro.The harvested hUCB-MSCs express the identical surface antigen as human bone marrow-derived mesenchymal stem cells(BM-MSCs).

Key words: human umbilical cord blood mesenchymal stem cells isolation culture