RA和bFGF体外诱导人牙髓干细胞向神经细胞分化的研究  

Induced differentiation of human dental pulp stem cells into neuron by RA and bFGF in vitro

作　　者：张智慧[1] 胡伟平[1] 郭阳[1] 曹潇方[1] 

机构地区：[1]哈尔滨医科大学附属第二医院口腔修复科,150086

出　　处：《实用口腔医学杂志》2011年第5期676-681,共6页Journal of Practical Stomatology

基　　金：黑龙江省自然科学基金(编号:D200856)

摘　　要：目的:探讨全反式维甲酸(all-trans-retinoic acid,RA)、碱性成纤维生长因子(basic fibroblast growth factor,bFGF)体外诱导人牙髓干细胞(dental pulp stem cells,DPSCs)分化为神经细胞的可行性。方法:体外分离培养DPSCs并进行克隆化,检测STRO-1的表达。将DPSCs分别接种于含有不同浓度RA、bFGF或二者结合的诱导液,MTT法检测细胞增殖能力,免疫荧光法检测微管相关蛋白(microtubule-associated protein-2,MAP-2)、神经元烯醇化酶(neuron-specific enolase,NSE)、胶质原纤维酸性蛋白(glial fibrillary acid protein,GFAP)的表达。透射电镜观察诱导前后细胞超微结构。结果:克隆来源细胞的STRO-1表达阳性。0.5μg/ml RA或20 ng/ml bFGF单独应用促增殖作用最强(P<0.05),bFGF单独作用各组及对照组均未检测出神经元样细胞。RA作用各组检测到阳性细胞。而0.5μg/ml RA与20 ng/ml bFGF联合应用的增殖和分化效应均优于其它组。透射电镜观察到幼稚神经元样细胞。结论:RA和bFGF联合应用可在体外有效诱导人DPSCs转化为神经细胞。Objective：To study the feasibility of all-trans-retinoic acid（RA） and basic fibroblast growth factor（bFGF） in the induction of human dental pulp stem cells（DPSCs） into neuron in vitro.Methods：DPSCs were in vitro cultured and cloned.Mesenchymal stem cell-specific markers（STRO-1） expression was detected.Cells were inoculated in different concentrations of the two factors alone or in combination for different times.MTT assay was used to examine the proliferation ability of the cells.The immune fluorescent assay was used to identify the expression of the microtubule-associated protein-2（MPA-2）,neuron-specific enolase（NSE） and glial fibrillary acidic protein（EFAP）,respectively.The ultrastructure of the cells was observed by TEM.Results：The expression of STRO-1 in cloned cells was positive.0.5 μg/ml RA or 20 ng/ml bFGF alone showed strong capacity for proliferation（P0.05）.The neuron-like cells were not detected in each group of bFGF and the control group.Positive cells were detected in the groups of RA.RA at 0.5 μg/ml combined with bFGF at 20 ng/ml showed strongest effects in the promotion of proliferation and differentiation of DPSCs.Conclusion：RA combined with bFGF may induce DPSCs into neuron-like and /or glia-like cells in vitro.

关 键 词：人牙髓干细胞 全反式维甲酸 成纤维生长因子-2 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]