RA和bFGF体外诱导人牙髓干细胞向神经细胞分化的研究

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。

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

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 assa...