神经干细胞原代培养及GABA能神经元的诱导分化  

Primary culture of neural stem cells and induction and differentiation of GABA neurons

作　　者：鲁明[1] 牛朝诗[1] 傅先明[1] 汪业汉[1] 钱若兵[1] 汤深凤[1] 王林[1] 李光武[2] 贾雪梅[3] 沈韶辉[3] 黄大可[3] 

机构地区：[1]安徽医科大学附属省立医院神经外科,安徽省立体定向神经外科研究所,合肥230001 [2]安徽医科大学神经生物学教研室,合肥230032 [3]安徽医科大学基础医学院形态中心,合肥230032

出　　处：《安徽医科大学学报》2006年第5期528-531,共4页Acta Universitatis Medicinalis Anhui

基　　金：安徽省高等学校自然科学研究项目(编号:2005KJ359zc)

摘　　要：目的 探讨神经干细胞（NSC）的原代培养及γ-氨基丁酸（GABA）能神经元的诱导分化、鉴定。方法 取出生1—3dSD大鼠全脑，分离、培养NSC并诱导分化了GABA能神经元，用免疫荧光染色进行鉴定。结果培养24h后，由单细胞发展成2～4细胞神经球，随时间延长球体增大，神经球均有Nestin阳性细胞；用含10％血清分化液分化1d后，神经球开始贴壁，伸出细长突起，部分可和周边神经球伸出的突起连接。神经球周边见大量散在贴壁的双极或多极细胞，免疫荧光染色可见NSE、GABA阳性细胞。加了分化液的实验组GABA阳性细胞分化率为（30．25±2．12）％，而对照组分化率为（1．14±．39）％。结论 成功从新生大鼠脑组织中分离、培养了NSC，并能较大比率分化GABA能神经元。Objective To investigate the primary culture of neural stem cells （NSCs） and induce differentiation of gamma amino butyric acid（GABA） neurons. Methods The whole brain of new born SD rat was taken and underwent primary culture. The NSCs were cultured, induced and differentiated into GABA neurons, which were identified using immunofluorescence stain. Results After 24 h of culture, neuropheres containing 2 to 4 cells appeared, and then they became bigger and bigger. Immunofluorescence stain found Nestin positive cells in the neuropheres. One day after administration of 10% blood differentiation serum, long synapse grew from the wall of the neurophere, which was able to connect to the synapses from the surrounding neuropheres. A great number of bipolar cells or multipolar appeared around the neuropheres. Immunofluorescence stain showed NSE, GABA, GFAP and Glu positive cells. The GABA positive cell differentiation rate was （30. 25 ± 2. 12）% in the experimental group, while it was （1.14 ± 1.39）% in the control group（P 〈 0. 01 ）. Conclusion The neural stem cells were isolated and cultured from new born SD rat using the present methods, which greatly increases the differentiation rate from NSCs to GABA.

关 键 词：干细胞/细胞学 神经形细胞学 细胞 培养 γ-氨 基丁酸 细胞分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R338.7[医药卫生—基础医学]