不同培养条件对神经干细胞分化的影响

Effects of different cultured conditions on differentiation of neural stem cells

目的:观察在不同血清浓度的诱导条件下神经干细胞定向分化为两类不同神经细胞的情况。方法:通过无血清培养的方法,将原代培养所获得的神经干细胞克隆经传代纯化后,分别接种在加入NeuralBasal,DMEM/F12+50g/L胎牛血清以及DMEM/F12+100g/L胎牛血清3种不同的培养基的培养皿中,7d后进行NF-200,GFAP免疫双标染色,在显微镜下随机选取20个视野,记数神经元和胶质细胞的比例。并进行统计学处理。结果:在无血清、50g/L以及100g/L胎牛血清浓度下,神经元与胶质细胞的比例分别为:65%:35%,45%:55%,33%:67%。在无血清和低血清浓度下,所分化的胶质细胞为原浆型星形胶质细胞,而在高血清浓度下则分化为纤维型星形胶质细胞。结论:血清浓度的高低决定了神经干细胞分化为神经元和胶质细胞的比例,并且对所分化的胶质细胞的种类也有影响。在无血清条件下神经干细胞趋向于向神经元分化。

AIM:To observe the differentiation of neural stem cells into two kinds of neural cells under inducing conditions of different serum concentration. METHODS:The neural stem cells purified from primary cultured clones by serum free media were placed into culture dishes of 3 different media: neural basal, DMEM/F12+50 g/L fetus bovine serum and DMEM/F12+100 g/L fetus bovine serum respectively. After 7 days, immunohistochemistry with antibody NF 200 and GFAP was taken to calculate the proportion of neurons and gliocyte by 20 fields of vision chosen randomly. The data were analyzed in statistics. RESULTS:The ratios of neuron and gliocytes in NB, DMEM+50 g/L fetus serum and DMEM+100 g/L fetus bovine serum were 65%:35%,45%:55%and 33%:67%respectively. The differentiated gliocytes in free and low serum media were protoplasmic astrocytes, and fibrous astrocytes in high fetus serum media. CONCLUSION:The concentration of serum decides the ratio of neural stem cells differentiation into neurons and gliocytes, and also influences the types of gliocytes it differentiates into.The neural stem cells have the tendency of differentiating into neurons under serum free medium.