原浆型星形胶质细胞调控神经干细胞定向分化的优化比例筛选

A selection of optimized ratio of the protoplasmic and fibrous astrocytes to the directional differentiation of neural stem cell

目的将神经干细胞(NSC)与原浆型星形胶质细胞(PAS)以不同比例进行体外共培养,观察NSC的定向分化的差异,筛选出有利于向神经元分化的最佳方案,为体内实验提供依据。方法根据NSC与PAS的比例进行分组:3:1组;2:1组;1:1组;1:2组,共培养10天后,采用免疫组化法对分化神经元进行DAPI荧光标记,在显微镜下随机选取20个视野,每次计数100个细胞,计算神经元在NSC中所占的比例,并进行统计学处理。结果NSC:PAS 2:1组分化得到的神经元数量最多,与分化的胶质细胞比例为65%:35%;而1:1组、3:1组稍低一些,比例分别为61%:39%和59%:41%;1:2组比例最低为57%:43%。结论在离体细胞共培养的实验中,因细胞的比例不同,NSC分化亦存在差异,其中NSC:PAS为2:1时,NSC分化为神经元的比例最高。

Objective Through culturing NSC with PAS in various ratios, we try to find the best ratio in which maximum amounts of neurons can differentiate from NSC. Methods By co-culturing of NSC with PAS in the ratio of 3 ： 1 or 2 ： 1 or 1 ： 1 or 1：2 for 10 days, immunohistocbemisty with DAPI was taken to calculate the proportion of neurons within 100 cells in 20 fields of vision chosen randomly. The data were analyzed statistically. Results The percentage of neurons differentiated from NSC in the ratio of 2：1 （65%） was much more than that in the other groups. The groups of 1：1 and 3：1 （61% and 59% ） were lower than the group of 2：1,the group of 1：2（57% ） was the lowest group. Conclusion In vitro co-culture of NSC and PAS in a ratio of 2 ： 1 may be more advantageous for NSC in differentiating into neurons.