人胎盘源间充质干细胞的分离、培养及生物学鉴定  

Isolation,Cultivation and Biological Identification of Human Placenta Mesenchymal Stem Cells

作　　者：赖平[1] 陈懿建[2] 罗耀玲[3] 张敏鸿[3] 杨建琼[3] 邱悦群[4] 

机构地区：[1]赣南医学院,江西赣州341000 [2]赣南医学院第一附属医院血液科,江西赣州341000 [3]赣南医学院第一附属医院临床科研中心,江西赣州341000 [4]赣南医学院第一附属医院,江西赣州341000

出　　处：《赣南医学院学报》2016年第2期183-186,共4页JOURNAL OF GANNAN MEDICAL UNIVERSITY

基　　金：江西省科技支撑计划项目(编号20141BBG70069)

摘　　要：目的:探讨机械剪碎组织加酶液消化法分离、培养人胎盘间充质干细胞的可行性,并对间充质干细胞进行生物学鉴定。方法:取足月产人胎盘,采用机械剪碎组织加酶液消化法分离,密度梯度离心法提纯胎盘组织中的细胞,利用细胞生长特性对细胞进行进一步分离、纯化并传代培养,倒置显微镜观察细胞形态,流式细胞术鉴定细胞表面标记。结果:在倒置显微镜下,细胞为贴壁生长,形态呈长梭形,少数为多角形,胎盘组织来源间充质干细胞在体外可稳定长期传代培养,应用流式细胞仪检测结果显示CD73、CD90、CD105表达阳性,CD34、CD45、HLADR表达阴性。结论:联合利用机械剪碎胎盘组织和酶消化法可高效获取胎盘间充质干细胞,获取的胎盘间充质干细胞在体外培养过程中生长稳定,增殖能力强,可长期传代或冻存。Objective： To investigate the feasibility of the combined use of mechanical minced placenta and enzymatic digestion to get and culture human placenta mesenchymal stem cells,and the biological identification of mesenchymal stem cells. Methods： Full-term human placenta was taken to seperate placental tissue cells by mechanical minced tissue joined with enzyme digestion. Placental tissue cells were purified by density gradient centrifugation,and further separated and purified with the growth characteristics of cells. Cells were observed under inverted microscope morphology and flow cytometry was adopted to identify cell surface markers. Results： Under inverted microscope,cells adherent growth,fusiform morphology,a small number of polygons were found. Placental tissues derived from mesenchymal stem cells in vitro could be long-term cultured stably. Flow cytometry showed that the expressions of CD73,CD90,CD105 were positive; the CD34,CD45,HLA-DR expressions were negative. Conclusion： Placenta derived mesenchymal stem cells can be efficiently obtained by the combined use of mechanical minced placenta and enzymatic digestion,can be cultured in vitro stably with high proliferative capacity,and can be long-term passed down or frozen.

关 键 词：人胎盘间充质干细胞 机械剪碎 酶消化 干细胞培养 

分 类 号：Q813.1[生物学—生物工程]