原代人脐带间充质干细胞培养方法的研究  

Culture methods for primary human umbilical cord mesenchymal stem cells

作　　者：王菲[1] 周洪[1] 郭昱成[1] 苏晓霞[1] 李昂[1] 赵小鹏 

机构地区：[1]西安交通大学口腔医院正畸科,陕西省西安市710004 [2]中国兵器工业第二零五研究所职工医院,陕西省西安市710065

出　　处：《中国组织工程研究》2014年第19期3042-3047,共6页Chinese Journal of Tissue Engineering Research

基　　金：陕西省自然科学基础研究计划(2014JM4146);陕西省"13115"科技创新工程计划项目(2008ZDKG-65)~~

摘　　要：背景:如何获得大量稳定活力好的细胞是人脐带间充质干细胞培养的难点。目的:通过组织块法、酶消化法和酶解组织块法3种细胞培养方法比较,筛选出最佳的人脐带间充质干细胞的培养方式。方法:分离新鲜脐带10根,分别采用组织块法、酶消化法、酶解组织块法3种方式培养人脐带间充质干细胞,比较3种方式原代人脐带间充质干细胞爬出所需时间、细胞培养成功率、绘制细胞增殖曲线,利用流式细胞仪检测细胞表面标志,并进行多项分化能力检测。结果与结论:酶解组织块培养法原代人脐带间充质干细胞爬出所需时间与酶消化法无显著差异,但明显短于组织块法(P<0.01),原代人脐带间充质干细胞培养成功率显著高于其他两组,人脐带间充质干细胞增殖情况3组无显著性差异,酶解组织块培养法培养的第3代人脐带间充质干细胞其细胞表面标志及多项分化能力符合间充质干细胞的特性。酶解组织块培养法能缩短原代人脐带间充质干细胞爬出时间,显著提高原代人脐带间充质干细胞的成功率。BACKGROUND：How to get a lot of stable and dynamic human umbilical cord mesenchymal stem cels is stil a difficulty. OBJECTIVE：To investigate the best culture way of human umbilical cord mesenchymal stem cels by comparing tissue explant, enzyme digestion, enzymolysis methods. METHODS：Ten fresh umbilical cords were isolated to human umbilical cord mesenchymal stem cels by using tissue explant, enzyme digestion, and enzymolysis methods, respectively. Hereafter, we compared cels harvested using three culture methods in terms of the time for the primary cels to creep, successful rates of cel-culturing, and curves of cellgrowth. Surface markers of cels were detected by flow cytometry and multiple differentiations of cels were detected. RESULTS AND CONCLUSION： The time for the primary cels to creep out in the enzymolysis and enzyme digestion groups had no significant difference, but both of them were significantly shorter than that in the tissue explant group （P 〈 0.01）. The successful rate of primary cellculture in the enzymolysis group was significantly higher than that in the other two groups. The cellproliferations of three groups had no significant difference. The cellsurface markers and differentiation ability of the third generation of human umbilical cord mesenchymal stem cels cultured by enzymolysis method met the characteristics of mesenchymal stem cels. In conclusion, the enzymolysis method can shorten the time for primary human umbilical cord mesenchymal stem cels to creep out and increase the successful rates of primary human umbilical cord mesenchymal stem cels.

关 键 词：干细胞 脐带脐血干细胞 间充质干细胞 人脐带间充质干细胞 原代细胞 细胞培养 组织块法 酶消化法 陕西省自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]