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背景:骨髓间充质干细胞(mesenchymalstemcells,MSCs)不仅能分化成间质细胞,而且能向实质细胞(如心肌细胞)转化。在体外定向诱导MSCs分化成神经细胞,是目前国内外研究热点,具有重大理论意义。目的:探讨大鼠MSCs体外诱导分化成神经元的能力。设计:随机空白对照实验的研究。地点、材料和干预:实验在吉林大学公共卫生学院毒理教研室完成。实验动物采用Wistar大鼠(吉林大学实验动物中心),6周龄,雌雄不限。将2～4代的rMSCs接种在铺有盖玻片的24孔板中。分为3个实验组,一个对照组。实验组分别加入终浓度为0.25,0.50和1.00mmol/L异丁基甲基黄嘌呤(isobutylmethylxanthine,IBMX)诱导传代的rMSCs,待细胞分化后进行形态学观察,对照组中不加诱导剂,并用免疫细胞化学方法进行细胞鉴定。主要观察指标:培养MSCs的生长情况,诱导后细胞的形态学变化及nestin(小鼠抗大鼠,单克隆),神经元特异性烯醇化酶(neuron-specificenolase,NSE)抗体(兔抗鼠,多抗)、微管相关蛋白-2a,b(microtubule-associatedprotein-2a,b,MAP-2a,b)的免疫细胞化学检测。结果:加入IBMX后,0.25mmol/L组分化成神经元样细胞较少。1.0mmol/L组细胞加入IBMX第2天开始可见细胞陆续死亡。0.5mmol/L组2d即可见有神经元样细胞出现,细胞具有典型的神经元形态特征,0
BACKGROUND: Mesenchymal stem cells (MSCs) not only differentiate into stromal cells, but also transform into parenchymal cells (such as cardiomyocytes). In vitro induced MSCs to differentiate into nerve cells, is the current research focus at home and abroad, has great theoretical significance. Objective: To investigate the ability of rat MSCs to differentiate into neurons in vitro. Design: A Randomized Blank Control Study. Location, Materials and Interventions: Experiments were performed at the Department of Toxicology, School of Public Health, Jilin University. Experimental animals Wistar rats (Jilin University Experimental Animal Center), 6 weeks old, male or female. 2 to 4 passages of rMSCs were plated in 24-well plates covered with coverslips. Divided into three experimental groups, a control group. The experimental group were respectively fed with rMSCs which were induced by isobutylmethylxanthine (IBMX) at the final concentrations of 0.25, 0.50 and 1.00mmol / L. After the cells were differentiated, the morphology was observed. In the control group, no inducer was added and immune cells Chemical methods for cell identification. MAIN OUTCOME MEASURES: The growth of MSCs was cultured, morphological changes of the cells were induced after the induction, and nestin (mouse anti-rat, monoclonal), neuron-specific enolase (NSE) Immunocytochemistry detection of microtubule-associated protein-2a, b, MAP-2a, b). Results: After addition of IBMX, 0.25mmol / L showed less differentiation into neuron-like cells. 1.0mmol / L group of cells added to IBMX 2 days began to see the cells died one after another. 0.5mmol / L group 2d can be seen neuron-like cells, the cells have a typical morphological characteristics of neurons, 0