大鼠骨髓间充质干细胞适宜分离和培养条件的研究  

Study on suitable isolation and culture of mesenchymal stem cell of rats in vitro

作　　者：吕铁伟[1] 田杰[1] 朱静[2] 刘官信[2] 

机构地区：[1]重庆医科大学儿科学院心内科 [2]重庆医科大学儿科研究所,重庆400014

出　　处：《重庆医科大学学报》2004年第6期715-717,721,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目 (项目编号 :3 0 2 713 81)。

摘　　要：目的 :寻求适宜有效的大鼠骨髓间充质干细胞 (mensechymalstemcell,MSCs)体外分离、纯化和培养的条件 ,为MSCs进一步的实验研究打下基础。方法 :分别用贴壁筛选法和密度梯度离心法来分离和纯化MSCs ,并用不同的接种密度、不同的血清浓度和不同的培养基检测细胞的生长情况 ,绘制出细胞的生长曲线 ;流式细胞仪检测MSCs生长周期。结果 :细胞贴壁呈纤维状生长 ,以 2× 10 9的浓度接种 ,在含 10 %胎牛血清的DMEM /F12培养基中 ,37℃、5 %CO2 细胞培养箱中 ,细胞生长状况良好 ;细胞在培养的最初几天处于生长的潜伏期 ,而后呈指数级生长 ,随着时间的延伸 ,细胞的生长速度减慢 ;而流式细胞仪检测示细胞大多处于G0 G1期。结论 :在适宜的培养条件下 ,MSCs在体外培养的过程中生长状态良好 ,增殖速度快 ,为进一步的研究打下了基础。Objective:To explore the method of isolation, purification and culture of rat mesenchymal stem cells (MSCs) in vitro in order to provide an experimental basis for further study.Methods:MSCs were acquired by adhesive screening and density gradient centrifugation methods.The growth of MSCs at different plate densities,different serum concentrations and different culture media were observed and their growth curves were charted.The cell cycle was detected by flow cytometer.Results:The adherent and fibroblast-shaped cells were in good growth state,which were plated at 2×10 9/ml and cultured in DMEM/F12 medium supplemented with 10% fetal bovine serum and placed in 37℃, 5%CO 2 incubator.Cells grew slowly the first several days after plated,then the growth accelerated at index speed.The results of the cell cycle detection demonstrated that most of MSCs were in G 0G 1 phase.Conclusion:In suitable conditions,MSCs show stable growth and rapid proliferation in vitro.

关 键 词：骨髓间充质干细胞 细胞培养 大鼠 

分 类 号：R331.22[医药卫生—人体生理学]