摘要
目的 :探讨从人脐带华通胶中分离培养间充质干细胞的方法,并对细胞进行标记,为细胞移植奠定基础。方法:采用组织块贴壁法分离培养人脐带华通胶间充质干细胞并进行传代,观察细胞的形态学特征,测定细胞生长曲线,通过流式细胞仪检测细胞免疫表型;用荧光染料PKH26对获取的细胞进行体外标记,观察标记细胞形态并记录其生长曲线。结果:组织块贴壁法可以成功地从人脐带华通胶中培养出间充质干细胞,并且在连续增殖传代过程中细胞形态未发生明显变化,细胞生长曲线相似;细胞能够高表达基质细胞抗原(CD29、CD90、CD44、CD105),而低表达造血干细胞抗原(CD34、CD45)和白细胞相关抗原(HLA-DR);PKH26可以标记脐带间充质干细胞,显示红色荧光,且标记细胞形态良好,与未标记细胞的生长曲线相似。结论:组织块贴壁法是一种简便有效的分离脐带间充质干细胞的方法;应用PKH26体外标记细胞成功率高、技术稳定。
Objective To explore the approach of separating, culturing and labeling mesenchymal stem cells （MSCs） from Wharton＇s jelly of human umbilical cord, so as to lay a foundation for the next step in cell transplantation. Methods By tissue explants adherent method, the MSCs were isolated and cultured from neonate umbilical cord, and then passaged. The morphological features of the cells were observed, and the cell growth curve was determined. The phenotype cell immunoreaetivity was detected by flow cytometry. The ceils were labeled with PKH26 in vitro. The morphology of the labeled cells was observed and the growth curve was recorded. Results MSCs were successfully obtained from Wharton＇s jelly of human umbilical cord via the proposed approach of tissue adherence. There were no significant changes of cell morphology occurred during the continuous proliferation and passage. The cell growth curve was similar. The flow cytometry analysis showed that cells highly expressed CD29, CD90, CD44 and CD105 of stromal cell antigen, while low expression occurred in hematopoietic stem cell associated antigens CD34, CD45 and leukocyte antigen HLA-DR. PKH26 could label umbilical cord mesenchymal stem ceils, and showed red fluorescence. The labeled cells were in good shape, and the growth curve was similar to that of the unlabeled cells. Conclusion Tissue block method is a simple and effective method for isolating mesenchymal stem cells from umbilical cord. The success rate of PKH26 labeled ceils in vitro is high, and the labeling technology is stable.
出处
《医疗卫生装备》
CAS
2016年第12期28-31,共4页
Chinese Medical Equipment Journal
基金
2015年度河北省医学科学研究重点项目(20150122)
关键词
脐带华通胶
间充质干细胞
细胞培养
细胞标记
细胞移植
umbilical cord
mesenchyma stem cell
cell culturing
cell labeling
cell transplantation
