摘要
目的探讨星形胶质细胞条件培养液(ACM)对神经干细胞(NSCs)体外分化的影响。方法行新生大鼠大脑皮质星形胶质细胞的纯化培养后收集ACM,将新生大鼠海马NSCs单克隆培养后行nestin免疫细胞荧光染色,诱导分化5d后行神经元特异性烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)、半乳糖脑苷脂(GALC)免疫细胞荧光染色;将单克隆培养的NSCs分为对照组和实验组,对照组为单纯NSCs培养液,实验组根据NSCs培养液与ACM比的不同分3组:A组(2:1),B组(1:1),C组(1:2)。各组培养1周,采用NSE免疫细胞荧光检测方法标记神经元并计数和统计学分析。结果纯化的星形胶质细胞胶质纤维酸性蛋白抗体标记阳性率为96.5%;单克隆培养的海马NSCs呈nestin阳性,诱导后呈NSE、GFAP和GALC阳性表达。ACM培养的海马NSCs各组分化为神经元的比例明显高于对照组(<0.01),其中A组的比例最高。结论ACM可以促进新生大鼠海马NSCs向神经元分化。
Objective To study the influence of astrocyte-conditioned medium （ ACM ） on the differentiation of neural stem cells （ NSCs ） in vitro. Methods Astrocytes from the cerebral cortex of neonate rats were cultured and purified, and ACM was collected. NSCs from hippocampus of neonate rats were obtained, immunofluoreseence stain was used to show nestin-, neuron specific enolase （ NSE ） -, glial fibrillary acidic protein （ GFAP ） - and galaeto cerebroside （ GALC ） -positive cells. Monoclone cultured NSCs were divided into control group （ NSCs medium ） and experimental groups （with different proportions of NSCs medium and ACM： A group 1：2, B group 1：1, C group 2：1）. The percentage of NSE positive cells was counted and the data was analyzed in statistics. Results The percentage of GFAP positive ceils was 96.5% in purified astrocytes, the cultured NSCs showed nestin positive, NSE, GFAP and GALC positive. The percentage of NSE positive cells differentiated from NSCs was much higher in experimental groups than in control group （P〈 0. 01）, with the highest percentage in A group. Conclusion ACM can promote the differentiation of NSCs into neurons in hiooocamous of neonate rats.
出处
《解剖科学进展》
CAS
2009年第3期265-268,共4页
Progress of Anatomical Sciences
基金
辽宁省自然科学基金(No.20082097)
