摘要
目的 研究不同浓度神经生长因子体外诱导大鼠骨髓间充质干细胞（BMSCs）向神经细胞分化的调节作用.方法 贴壁培养BMSCs,流式细胞检测表面标志物.A组加入3种浓度EGF和bFGF;B组加同样浓度bFGF;C组不加诱导剂.A、B组用脑源性神经营养因子（BDNF）和全反式维甲酸（ATRA）诱导,免疫荧光及免疫组化鉴定.结果 第3代BMSCs为梭形样细胞,CD29、CD44、CD90为阳性,CD34、CD45为阴性.A组中100μg/L巢蛋白（NESTIN）（81.9±1.8）%.A组与B组比较,差异有统计学意义（P〈0.05）.继续诱导,可见微管相关蛋白2（MAP2）、神经丝（NF）及神经元特异性烯醇化酶（NSE）阳性细胞.结论 采用改良的全骨髓贴壁法,细胞活性好.bFGF和EGF同时应用比bFGF单独应用效果好.100μg//L（bFGF＋EGF）预诱导m神经干细胞数量多,并分化为神经元样细胞.
Objective To investigate the effect of BMSCs induced by nerve growth factor on different concentrations. Methods BMSCs were cultured with adherent culture and surface markers were identifed by flow cytometry. BMSCs in Group A were added three kinds of EGF ＋ bFGF; BMSCs in Group B were treated bFGF with the same conceration; BMSCs in Group B were processed without inducer. BMSCs in both Group A and Group B were induced by brain derived neurotrophic factor （BDNF） and retinoic acid （ATRA） and then the results were identified by immunofluorescence and Immunohistochemcical staining. Results The third generation of BMSCs appeared spindle-shaped cells and the expression of CD29, CD44 and CD90 was positive and the expression of CD34 and CD54 was negative. The expression of 100 ug/L Nestin in BMSCs was high （81.9 ±1.8）% in Group A which had statistical differences compared with that in Group B （ P 〈 0.05）. After continuing induce, the BMSCs expression of MAP2, NF and NSE in BMSCs was possitive. Conclusion This study utilizes and modifies total bone marrow adherence with good cytoactive. The better effect is acquired with the combination of bFGF and EGF than the bFGF only. A great deal of BMSCs can be pre-induced by 100 ug/L （bFGF ＋ EGF） which can be differentiated into nerve stems neuron-like cells.
出处
《中国美容整形外科杂志》
CAS
2010年第8期510-512,共3页
Chinese Journal of Aesthetic and Plastic Surgery
基金
基金项目：辽宁省教育厅高校科技攻关基金资助项目（1081225-1-00）
关键词
骨髓间充质干细胞
神经生长因子
神经元样细胞
诱导
分化
Bone marrow stromal mesertchymal stem cells
Nerve growth factor
Neuron-like cell
Induction
Differentiation.
