中国组织工程研究与临床康复2011，Vol.15Issue(23)：4220-4224,5.DOI:10.3969/j.issn.1673-8225.2011.23.010
人脐带来源间充质干细胞分离培养方法的优化
Optimized study on isolation and cultivation of human umbilical cord mesenchymal stem cells
摘要
Abstract
BACKGROUND: There are different methods to isolate and culture human umbilical cord mesenchymal stem cells (hUC-MSCs), so how to fast and efficiently harvest hUC-MSCs have become a research hotspot.OBJECTIVE: To optimize digestive enzymes components for the preparation of hUC-MSCs, in order to lay the foundation of preparation for hUC-MSCs.METHODS: Human umbilical cords were collected from full term deliveries under aseptic conditions. According to different mixed enzyme concentration ratio, the samples were divided into three groups: mixed enzyme Ⅰ group, mixed enzyme Ⅱ group, mixed enzyme Ⅲ group. Then according to digest time each group into three subgroups: 1 hour, 2 hours, and 3 hours. Finally, suspended cell volume was decided as 4 mL to count cells. Dulbecco’s Modified Eagle Medium with fetal bovine serum was used for cell culture.RESULTS AND CONCLUSION: Based on the three digestion enzyme concentration, at the duration of 1 hour, 2 hours and 3 hours, mixed enzyme Ⅲ group had the highest total cell number and the total cell rate was statistically different from the other groups (P ＜ 0.05). At the duration of 3 hours, live cell rate was the lowest in the mixed enzyme Ⅲ group, and there was statistically significant differences among these groups (P ＜ 0.01). The optimal isolation for hUC-MSCs is 0.3％ collagenase Ⅱ, 0.1％ trypsin, 0.02％ EDTA, 0.1％ hyaluronidase and 0.1％ DNA enzymeⅠ digestion for 2 hours.关键词
脐带/间充质干细胞/Ⅱ型胶原酶/透明质酸酶/DNA酶Ⅰ分类
医药卫生引用本文复制引用
齐凯,董丽媛,陈显久,赵婕,薛国芳,陈彦,牛勃..人脐带来源间充质干细胞分离培养方法的优化[J].中国组织工程研究与临床康复,2011,15(23):4220-4224,5.基金项目
国家自然科学基金(30600226,30472251). （30600226,30472251）
