CAJ | 학술논문

背景：从体外分离培养出高纯度、生物学性能均一的神经干细胞，建立起一套完整的神经干细胞培养体系，是进行神经干细胞研究的基础。 　　目的：建立新生小鼠海马、嗅球、皮质组织神经干细胞的分离培养体系，并对其生物学特性进行分析。 　　方法：分离新生昆明小鼠海马、嗅球、皮质组织，采用机械分离和胰酶消化法提取原代神经干细胞。采用无血清培养技术、机械吹打和酶消化法进行传代培养神经干细胞。以体积分数为10%的胎牛血清诱导分化神经干细胞。对神经干细胞及其分化产物行CD133、巢蛋白、β-微管蛋白Ⅲ、胶质纤维酸性蛋白免疫荧光染色鉴定。 　　结果与结论：从新生小鼠海马、嗅球、皮质可提取出具有自我更新和多向分化能力的神经干细胞，经巢蛋白、CD133免疫荧光染色检测呈阳性；神经干细胞经胎牛血清诱导后可分化为β-微管蛋白Ⅲ、胶质纤维酸性蛋白阳性细胞，并证实染色阳性细胞为神经元和星形胶质细胞。该实验建立了一套神经干细胞体外分离培养、纯化、鉴定、诱导分化方案，为后续神经干细胞研究的顺利进行奠定了实验基础。
배경：종체외분리배양출고순도、생물학성능균일적신경간세포，건립기일투완정적신경간세포배양체계，시진행신경간세포연구적기출。 　　목적：건립신생소서해마、후구、피질조직신경간세포적분리배양체계，병대기생물학특성진행분석。 　　방법：분리신생곤명소서해마、후구、피질조직，채용궤계분리화이매소화법제취원대신경간세포。채용무혈청배양기술、궤계취타화매소화법진행전대배양신경간세포。이체적분수위10%적태우혈청유도분화신경간세포。대신경간세포급기분화산물행CD133、소단백、β-미관단백Ⅲ、효질섬유산성단백면역형광염색감정。 　　결과여결론：종신생소서해마、후구、피질가제취출구유자아경신화다향분화능력적신경간세포，경소단백、CD133면역형광염색검측정양성；신경간세포경태우혈청유도후가분화위β-미관단백Ⅲ、효질섬유산성단백양성세포，병증실염색양성세포위신경원화성형효질세포。해실험건립료일투신경간세포체외분리배양、순화、감정、유도분화방안，위후속신경간세포연구적순리진행전정료실험기출。
BACKGROUND:To in vitro isolate neural stem cel s with high purity and uniform biological properties and to establish a complete set of neural stem cel culture system is the basis for neural stem cel research. OBJECTIVE:To establish an isolation and culture system for neural stem cel s from newborn mouse hippocampus, olfactory bulb and cortex and to analyze the biological properties of cel s. METHODS:Neural stem cel s were isolated from the hippocampus, olfactory bulb and cortex tissue of newborn Kunming mice by mechanical separation and trypsin digestion. Serum-free culture technology, mechanical pipetting and trypsin digestion were used for subculture of neural stem cel s. 10%fetal bovine serum was used to induce differentiation of neural stem cel s. Neural stem cel s and their differentiated products were identified by immunofluorescent staining of Nestin, CD133,β-TubulinIII, glial fibril ary acidic protein. RESULTS AND CONCLUSION:The neural stem cel obtained from newborn mouse hippocampus, olfactory bulb and cortex had the capacity of self-renewal and differentiation which were positive for Nestin and CD133. After induction with fetal bovine serum, neural stem cel could differentiation toβ-tubulinIII or glial fibril ary acidic protein positive cel s that were neurons and astrocytes. This experiment has successful y established the neural stem cel isolation, culture, identification and induction system, providing experimental basis for subsequent studies of neural stem cel s.