基础医学与临床 ›› 2009, Vol. 29 ›› Issue (6): 566-569.

• 研究论文 • 上一篇    下一篇

绿色荧光蛋白转基因小鼠骨髓基质细胞体外诱导分化为神经元样细胞

周辉 姜晓丹 徐如祥 蔡颖谦 邹雨汐   

  1. 南方医科大学珠江医院 南方医科大学珠江医院
  • 收稿日期:2008-09-16 修回日期:2008-10-30 出版日期:2009-06-25 发布日期:2009-06-25
  • 通讯作者: 姜晓丹

Bone marrow stromal cells of GFP transgenic mice differentiate into neuron-like cells in vitro

Hui ZHOU, Xiao-dan JIANG, Ru-xiang XU, Ying-qian CAI, Yu-xi ZOU   

  1. Zhujiang Hospital of Southern Medical University Zhujiang Hospital of Southern Medical University
  • Received:2008-09-16 Revised:2008-10-30 Online:2009-06-25 Published:2009-06-25
  • Contact: Xiao-dan JIANG,

摘要: 目的 研究绿色荧光蛋白转基因小鼠骨髓基质细胞(GFP-GM-BMSCs)在体外无血清培养基+神经细胞因子诱导条件下,向神经细胞分化的能力。方法 用贴壁法体外培养GFP-GM-BMSCs,取第3代GFP-GM-BMSCs,用含浓度均为20μg/L的表皮生长因子(EGF)和碱性成纤维细胞生长因子(bFGF)的无血清培养基(neurobasal-A+2%B27)诱导分化。第5天用免疫细胞荧光方法检测巢蛋白(nestin)的表达,第10天用神经元烯醇化酶 (NSE)、神经胶质纤维酸性蛋白(GFAP)免疫细胞化学方法鉴定阳性细胞。结果 GFP-GM-BMSCs经无血清培养基+神经细胞因子诱导后,细胞胞体变圆,伸出细长突起, 有的突起连接成网,呈神经元样形态。诱导第5天,nestin阳性表达的细胞为40.24%+5.09%;第10天,NSE阳性、GFAP阳性的细胞分别为36.43%+5.27%和49.73%+6.28%。 结论 GFP-GM-BMSCs在体外含EGF、bFGF的无血清培养基中,能分化成神经元样细胞。

关键词: 绿色荧光蛋白, 骨髓基质细胞, 细胞分化, 神经元样细胞

Abstract: Objective To induce bone marrow stromal cells (BMSCs) of green fluorescent protein (GFP) transgenic mice (GFP-GM-BMSCs) into neural-like cells in serum-free medium. Methods GFP-GM-BMSCs were isolated and cultured in serum-containing medium. Then the 3rd generation of GFP-GM-BMSCs were transferred into serum-free medium containing 2% B27 supplement with 20ng/ml basic fibroblast growth factor (bFGF) and 20ng/ml epidermal growth factor (EGF).At the 5th day after culture, the expression of Nestin was examined by immunofluorescence. NSE, GFAP were examined by immunocytochemistry on the 10th day following the induction. Results The induced GFP-GM-BMSCs gradually became circular. Some of them grew long processes and showed the neuron-like appearance. The positive expression of Nestin was about (40.24%+5.09)% on the 5th day, while the NSE and GFAP were positively expressed as (36.43+5.27)% and (49.73+6.28)% respectively on the 10th day after the culture. Conclusions GFP-GM-BMSCs could be induced into neuron-like cells in serum-free medium containing EGF and bFGF.

Key words: green fluorescent protein (GFP), bone marrow stromal cells (BMSCs), cell differentiate, neuron-like cell