目的：观察改良型生物硅胶膜对神经干细胞（NSCs）增殖与分化的影响，为选择NSCs培养载体提供依据。方法选取孕16 d胎鼠的大脑皮质作为细胞来源，建立胚胎NSCs的体外培养体系并随机分为观察组和对照组，其中观察组接种于改良型生物硅胶膜上、对照组接种于普通培养皿中。普通倒置显微镜及电镜下观察细胞生长发育形态学变化，免疫荧光染色技术对NSCs及诱导分化细胞进行鉴定，四甲基偶氮唑蓝（ MTT）法检测细胞增殖情况。结果培养2 d后，显微镜下两组均可见神经细胞球生成，贴壁良好，细胞球边缘有突起呈辐射状向外发出，神经球NSCs特异标志物神经巢蛋白均呈阳性；培养7 d后，观察组和对照组贴壁分化的神经元烯醇化酶阳性细胞率分别为25．14％±1．98％、24．67％±2．05％，胶质纤维酸性蛋白阳性细胞率分别为59．48％±2．07％、60．87％±1．08％，组间比较，P均＞0．05；两组生长曲线亦无显著差异（P＞0．05）。结论改良型生物硅胶膜对NSCs的生长、增殖及诱导分化无明显影响，但能促进细胞之间突起的增长，可作为培养载体用于研究力学因素对NSCs增殖、诱导分化的影响。%Objective To explore the influence of silicone membrane on the neural stem cells'( NSCs) proliferation and differentiation, and to provide an experimental basis for the research of NSCs'carrier.Methods We selected 16 d pregnant rats cerebral cortex as the source of the cells , and established embryonic neural stem cells (NSCs) culture system in vitro. Then NSCs were randomly divided into the observation group and the control group , the observation group was inoculated on modified silicone membrane , and the control group was inoculated on the ordinary culture dish .The cell growth morphology of NSCs was observed by using light microscope and electron microscope .The NSCs and differentiated cells were identified by immunofluorescent staining , and MTT assay was used to detect the proliferation .Results After culture for 2 d, neural spheres were formed and adhered well , neuritis was detected from the assay , and nestin-positive was detected in both groups . After 7 d, NSE-positive rates of the experimental group and control group were 25.14%±1.98%and 24.67%±2.05%;GFAP-positive 59.48%±2.07%and 60.87%±1.08%(all P>0.05).The growth curve of the two groups showed no sig-nificant difference (P>0.05).Conclusions The modified silicone membrane has no significant effect on the NSCs'growth, proliferation and induced differentiation but promotes the growth of neurite between cells , which can be used as a culture car-rier to study the effects of mechanical factors on NSCs'proliferation and differentiation .
展开▼
