【摘要】 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。
【关键词】 星形胶质细胞;神经干细胞;分化;比例
A selection of optimized ratio of the protoplasmic and fibrous astrocytes to the directional differentiation of neural stem cell
LI Min,ZEGN Lin,LIU Yuan,et al.
(State Key Laboratory of Trauma,Burns and Combined Injury,Department 3,Institute of Surgery Research,Daping Hospital,Third Military Medical University,Chongqing 400042,China)
Abstract: 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 coculturing of NSC with PAS in the ratio of 3:1 or 2:1 or 1:1 or 1:2 for 10 days, immunohistochemisty 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 coculture of NSC and PAS in a ratio of 2:1 may be more advantageous for NSC in differentiating into neurons.
Key words:astrocyte;neural stem cell;differentiation;ratio
本室在实验中观察到原浆型星型胶质细胞(protoplasmic astrocyte，PAS)与纤维型星型胶质细胞(fibrous astrocyte，FAS)相比更有利于神经干细胞(neural stem cell，NSC)向神经元的分化[1]。但不同比例条件下NSC向神经元分化的情况可能存在差异，为此本实验将NSC与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。
材料与方法
1 实验动物
孕13～14天的大鼠和新生2天的大鼠。
2 试剂与材料
10%胎牛血清(FB
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