小鼠海马神经元细胞系HT22微环境诱导BMSCs神经分化  

Micro-environment of murine hippocampus neurons cell line HT22 induced the differentiation of BMSCs into neurons

作　　者：侯伟健[1] 王昕[2] 郝世宇 佟浩[1] 敖强[1] 王佐周[3] 柏树令[1] 

机构地区：[1]中国医科大学公共基础学院组织工程教研室,辽宁沈阳110122 [2]中国医科大学生物医学工程系,辽宁沈阳110122 [3]中国医科大学基础医学院病理教研室,辽宁沈阳110122

出　　处：《解剖科学进展》2016年第4期411-415,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金(30940095);辽宁省教育厅基金(L2010575);辽宁省科技厅社发项目(201222 5016)

摘　　要：目的探讨小鼠海马神经元细胞系HT22的微环境对骨髓间充质干细胞(BMSCs)分化的定向诱导作用。方法分别培养HT22细胞和GFP转基因荧光小鼠的BMSCs;直接接触法和间接接触法共培养,荧光显微比较GFP标记的干细胞形态变化;收集间接接触共培养的BMSCs行Western blot检测神经相关的蛋白(GFAP,NSE)的表达和变化;利用免疫荧光法检查GFAP和NSE抗原在细胞内的表达。结果培养的骨髓贴壁细胞为CD11b、CD45阴性,CD90阳性。与HT22细胞直接接触共培养及间接接触共培养的BMSCs细胞长轴增长,轴突样突起明显;经HT22细胞培养液诱导的BMSCs内GFAP、NSE表达水平随时间增加而增强;与HT22细胞直接接触共培养诱导的BMSCs表达GFAP和NSE。结论小鼠海马神经元细胞系HT22的微环境可诱导BMSCs向神经元样细胞分化。Objective To study the effect of murine hippocampus neurons cell line HT22 micro-environment on the differentiation of bone marrow mesenchymal stem cells（BMSCs） into neurons. Methods HT22 cells and BMSCs derived from fluorescence GFP transgenic mice were co-cultured with direct contact or indirect contact. The mophology of BMSCs was observed under fluorescence microscopy, the expression and localization of GFAP and NSE in BMSCs were detected by Western blot and immunofluorescence. Results CD90 positive BMSCs were observed, but negative for CD1 lb or CD45 by flow cytometry. The long processes of BMSCs were observed in ether diect or indiect coulmre with H322, with axons-like protuberance obviously. The expression levds of GFAP and NSE proteins we, re, increased in direct contaet cocuhured BMSCs with H322 cells as culture time prolonged. Conclusion The microenvironment of the hippocampus neuron Ht22 cells in mice induced the differentiation of BMSCs into neuron-like ceils.

关 键 词：小鼠海马神经元 HT22细胞 骨髓间充质干细胞 神经分化 微环境 

分 类 号：Q254[生物学—细胞生物学]