摘要
建立人脐带间充质干细胞标准化的制备工艺.比较不同培养基、消化液及消化时间处理人脐带间充质干细胞的增殖水平、表型、分化功能;设计不同细胞接种密度及汇合度进行培养,观察细胞生长状态并计算细胞倍增时间.人脐带间充质干细胞在MSC-T4培养基中,CTS消化酶消化3 min表现出更佳的增殖水平、间充质干细胞表型、成骨、成脂的能力;10 000 cells/cm2和8 000 cells/cm2的细胞接种密度分别是最合适的人脐带间充质干细胞复苏接种密度和传代接种密度;90％～95％汇合度为人脐带间充质干细胞传代最佳汇合度.
Abstract
Human umbilical cord mesenchymal stem cells have shown varying degrees of efficacy in the treatment of a variety of refractory diseases,and the quality of stem cells is an important guarantee of efficacy.At present,different laboratories have different methods and key parameters for the preparation of stem cells,resulting in different quality of prepared stem cells.How to establish a set of scientific,stable and high-quality human umbilical cord mesenchymal stem cell preparation process is the goal of stem cell preparation engineering team.To establish a standardized preparation process for human umbilical cord mesenchymal stem cells(hUC-MSCs).Different culture media,digestion solutions,and digestion times were compared to evaluate the proliferation level,phenotype,and differentiation potential of hUC-MSCs.Different cell seeding densities and confluence rates were designed for cultivation,and the cell growth status and doubling time were observed.hUC-MSCs showed better proliferation level,phenotype,osteogenic,and adipogenic abilities when cultured in MSC-T4 medium with a 3-minute digestion using CTS enzyme.The optimal cell recovery seeding density and passage seeding density for hUC-MSCs were determined to be 10 000 cells/cm2 and 8 000 cells/cm2,respectively.A confluence rate of 90％～95％was found to be optimal for hUC-MSCs passage.
基金项目
佛山市科技局科技计划项目(2320001009209)
NETL
NSTL
万方数据