人胎盘底蜕膜间充质干细胞的分离培养及其多向分化潜能的实验研究  

Isolation and multipotent differentiation of human decidua basalis-derived mesenchymal stem cells

作　　者：卢国辉[1] 张世忠[2] 陈强[2] 王雪峰[3] 卢凤飞[2] 刘剑[2] 李明[2] 李振勇[2] 

机构地区：[1]南方医科大学珠江医院神经外科研究所2妇产科,广东广州510282 [2]南方医科大学珠江医院神经外科研究所,广东广州510282 [3]南方医科大学珠江医院妇产科,广东广州510282

出　　处：《南方医科大学学报》2011年第2期262-265,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(30772232);广东省科技计划项目(2009B030801375)~~

摘　　要：目的探讨人胎盘底蜕膜间充质干细胞(MSCs)体外生物学特性,并验证其多向分化潜能。方法通过酶消化和密度梯度离心法从人足月胎盘底蜕膜中获取MSCs,倒置显微镜下观察其形态变化和生长特点;CCK-8试剂盒检测细胞增殖能力;流式细胞仪检测细胞周期及表面抗原(CD29、CD44、CD73、CD90、CD34、CD45、CD14)的表达;在特定诱导条件下,使其向成骨、成脂、成软骨方向分化,鉴定其多向分化潜能。结果人足月胎盘底蜕膜中分离的MSCs呈克隆样生长,形态类似于骨髓MSCs,传代后细胞增殖迅速,细胞倍增时间为(2.21±0.21)d;大于70%的细胞处于静止期(G0/G1期),阳性表达CD29、CD44、CD73、CD90,不表达CD34、CD45、CD14;经诱导后茜素红染色、油红O染色和Alcian blue染色均呈阳性表现。结论胎盘底蜕膜中含有丰富的MSCs,易于体外分离培养,且具有多向分化潜能,有希望成为另一新颖的干细胞来源。Objective To investigate the biological features of human decidua basalis-derived mesenchymal stem cells（PDB-MSCs） in vitro and identify their capacity of multilineage differentiation.Methods PDB-MSCs were harvested from the decidua basalis of term placental by enzymatic digestion and density gradient centrifugation,and the growth characteristics and morphological changes of the MSCs were observed by inverted microscope.The proliferative ability of the cells was assessed by Cell Counting Kit-8.The cell cycle and expressions of the surface markers（CD29,CD44,CD73,CD90,CD34,CD45,and CD14） of the MSCs were identified by flow cytometry.Multilineage differentiation capacity of the cells was tested by inducing their differentiation toward osteoblasts,adipocytes and chondroblasts in vitro.Results MSCs isolated from human decidua basalis of term placental exhibited a morphology similar to that of bone marrow-derived MSCs,and grew into colonies in in vitro culture,where the cells proliferated rapidly after passage with a cell doubling time of 2.21±0.21 days.More than 70% of the cells stayed in the resting stage（G0/G1） and showed positivity for CD29,CD44,CD73 and CD90,but not for CD14,CD34 or CD45.After induction,the cells showed positive results of alizarin red staining,oil red O staining and Alcian blue staining.Conclusion Human decidua basalis contains a rich source of MSCs,which can be easily isolated and cultured without affecting their capacity of multilineage differentiation.The PDB-MSCs may have the potential as a novel source of stem cells.

关 键 词：间充质干细胞 底蜕膜 分离培养 多向分化 

分 类 号：R331.22[医药卫生—人体生理学] R329.24[医药卫生—基础医学]