大鼠骨髓间充质干细胞的培养及鉴定
Culture and identification of rat marrow mesenchymal stem cells
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摘要: 目的: 分离培养和鉴定大鼠骨髓间充质干细胞。方法: 分离SD青年大鼠股骨,L-DMEM冲洗骨髓腔,制备细胞悬液接种培养,消化传代纯化细胞。传代的骨髓间充质干细胞进行增殖曲线、克隆形成能力测定,用免疫组化检测增殖细胞vimentin、laminin的表达。结果: 传代的细胞在培养第1~2天为潜伏期,第3~6天是细胞快速增殖期,第6天达高峰,第9天后速度减慢,进入平台期,并且随传代次数的增加,细胞增殖速度逐渐下降。随传代次数增多,克隆形成能力逐渐下降。第5代以后细胞基本纯化为骨髓间充质干细胞,细胞呈长梭形。免疫组化检测增殖细胞vimentin表达阳性、laminin表达阴性。结论: 本实验方法能有效扩增大鼠骨髓间充质干细胞,而且简单易行。Abstract: Objective: To culture and identify rat mesenchymal stem cells(MSCs).Methods: Rat MSCs were separated from femur marrow by flushing out with low glucose DMEM,and planted in plastic culture flask.MSCs were observed by means of growth curve and clone forming rates.Vimentin,laminin were detected by immunohistochemistry.Results: The growth curve of MSCs of passage culture as follows:latent period 1-2 days;logarithm period 3-6 days and platform period afterward.Clone forming rates decreased with passage.The cells were purified after fifth passage of MSCs and shown as spindle-shaped morphology.Conclusions: The cultured cells are rat MSCs.The method is easy to operate and simple.
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