论文部分内容阅读
目的比较组织块贴壁培养法、脐带匀浆胶原酶消化法与改良胶原酶消化法制备脐带间充质干细胞(CMSCs),探讨一种简便、快速、高质量的CMSCs制备方法。方法无菌条件下留取健康孕妇剖宫产的新生儿脐带,采用上述三种方法获取CMSCs,经传代培养、倒置显微镜形态观察、细胞增殖能力观察及流式细胞仪检测CMSCs免疫表型。结果三种方法均制备出CMSCs,但改良胶原酶消化法贴壁迅速,短时间内即可获得大量的CMSCs。细胞免疫表型:高表达CD90、CD105、CD73、CD29、CD44;极低表达CD45、CD34、CD14、CD19、HLA-DR。结论改良胶原酶消化法可以快速分离脐带华尔通胶(Wharton’sjelly)中CMSCs,且细胞迅速贴壁生长3～5 d即可传代,是一种简便快速易行的CMSCs制备方法。
OBJECTIVE: To compare the method of adherent culture of tissue explants, umbilical cord plasminogen digestion and modified collagenase digestion to prepare cord blood mesenchymal stem cells (CMSCs), and to explore a simple, rapid and high quality method for preparing CMSCs. Methods Umbilical cord of cesarean section was collected from healthy pregnant women under aseptic conditions. The CMSCs were obtained by the above three methods. The subcultured CMSCs were observed under inverted microscope, the proliferation of CMSCs was observed and the immunophenotypes of CMSCs were detected by flow cytometry. Results CMSCs were prepared by all three methods. However, modified collagenase digestion method could rapidly attach to CMS CMS in a short period of time. Cell immunophenotype: high expression of CD90, CD105, CD73, CD29, CD44; very low expression of CD45, CD34, CD14, CD19, HLA-DR. Conclusion The modified collagenase digestion method can rapidly isolate CMSCs from umbilical cord blood Wharton ’s jelly, and the cells can be passaged rapidly by adherent growth for 3 to 5 days. It is a simple and rapid method for preparing CMSCs.