骨髓间充质干细胞体外分化过程中细胞周期蛋白的表达变化  

Expression changes of cyclin in in-vitro differentiation of bone mesenchymal stem cells

作　　者：吕翠[1] 付文玉[1] 刘倩[1] 潘顺[1] 季泰令[1] 曾现伟[1] 

机构地区：[1]潍坊医学院附属医院干细胞研究所,山东潍坊261031

出　　处：《中国卫生检验杂志》2012年第1期5-7,共3页Chinese Journal of Health Laboratory Technology

基　　金：山东省科技攻关计划资助项目(2011GSF11829)

摘　　要：目的:研究大鼠骨髓间充质干细胞(MSCs)体外定向分化为多巴胺(DA)能神经元过程中细胞周期相关蛋白的表达变化。方法:体外分离、扩增和鉴定大鼠骨髓MSCs。碱性成纤维细胞生长因子(bFGF)预诱导24 h后,用1μmol/L全反式维甲酸(ATRA)和50 ng/ml GDNF联合诱导6 d,检测神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)、酪氨酸羟化酶(TH)、增殖细胞核抗原(PCNA)、Bcl-2、细胞周期蛋白依赖性激酶(CDK2)、细胞周期蛋白(Cyclin)B1的表达。结果:诱导后大鼠骨髓MSCs能分化为具有典型神经元形态的细胞,大鼠MSCs经诱导后PCNA、CDK2、Cyclin B1的表达均较对照组明显减弱,Bcl-2表达与对照组无明显差别。结论:骨髓间充质干细胞体外开始分化后细胞增殖力下降。Objective：To explore the expressions of cell cycle associated proteins in the process of rat bone marrow MSCs differentiating into dopaminergic neurons in vitro. Methods： The MSCs derived from rat bone marrow were cultured and passaged in vitro. After being induced by basic fibroblast growth factor（bFGF） for 24 h, the pured MSCs of the third generation were induced into dopaminergic neurons by using 1 μmol/L all - trans retinoic acid （ATRA） and 50 ng/ml glial - derived neurotrophic factor （GDNF） for 6 d. Then the expressions of neuron specific enolase（ NSE）, glia fibrillary acid protein（ GFAP）, tyrosine hydroxylase （ TH）, proliferating cell nuclear antigen （ PCNA）, Bcl - 2, cyclin - dependent protein kinase - 2 （ CDK2 ） and CyclinB 1 were detected. Results： Under the inverted microscope, the induced MSCs showed typical neuronal morphological characteristics after induction. Compared with the MSCs in control, the expressions of PCNA, CDK2 and Cyclin B1 of the induced cells down - regulated significantly, and the expression of Bcl -2 had no significant difference. Conclusion： Induced MSCs slowed their prolifera- tion and down regulated cell cycle activators as they differentiated.

关 键 词：间充质干细胞 多巴胺能神经元 分化 细胞周期相关蛋白 大鼠 

分 类 号：Q462[生物学—生理学]