大鼠骨髓间质干细胞体外培养扩增及生物学特性研究  

Study on Method of Expanding and Culturing Rat Mesenchymal Stem Cells in vitro and Its Biological Characteristics

作　　者：余勤[1] 连俊兰[1] 王艳[1] 郭莹[1] 万海同[1] 

机构地区：[1]浙江中医药大学,杭州310053

出　　处：《浙江中医药大学学报》2006年第2期189-192,共4页Journal of Zhejiang Chinese Medical University

基　　金：国家自然科学基金资助计划项目(No:30371789);浙江省科技厅资助项目(No:2004C33014)~~

摘　　要：[目的]建立从大鼠骨髓中分离大鼠骨髓间质干细胞（rat mesenchymal stem cells，rMSCs）和体外传代培养技术，并对rMSCs的生物学特性进行研究。[方法]采用贴壁筛选法分离rMSCs，并通过不断传代进行纯化和扩增培养，绘制1、3、5代细胞生长曲线，用流式细胞仪检测其表面抗原，并比较在含不同浓度胎牛血清培养液培养后rMSCs的生长、增殖情况。[结果]rMSCs在体外培养扩增，原代可获得（5～6）×10^5、第5代可获得（2～3）×10^8个细胞。rMSCs形态呈长梭形，细胞生长曲线呈S形，流式细胞仪检测结果显示，rMSCs表面抗原CD90表达阳性，而CD45表达阴性。[结论]所建立的分离和培养方法获取的是rMSCs，具有rMSCs生物学特性，是组织工程研究中良好的细胞来源。[Objective] To establish a method to seperate rat mesenchymal stem cells（rMSCs） from rat marrow, culture rMSCs in vitro, and study its biological characteristics. [Methods] rMSCs were seperated from rat marrow with wall-sticking method and expanding cultured in vitro. To draw the growth curves of cells at 1th, 3th, 5th passage, detect the surface antigens with flow cytometry. To evaluate the effect of fetal calf serum with different concentrations to rMSCs. [Results] There were approximately （5 -6） ×10^5 cells after primary culture and over （2-3））×10^8 rMSCs at 5th passage. The appearance of rMSCs is like shape of spear and its growth curves are like shape of S. The rMSCs were positive for CD90 and negative for CD45. [Conclusions]Cells seperated and cultured by the method established in this experiment were rMSCs. They have the biological characteristics of rMSCs, and are good resources for study in tissue engineering.

关 键 词：间质干细胞 骨髓 生物学特性 大鼠 

分 类 号：R392.2[医药卫生—免疫学]