摘要
目的 探讨体外分离和培养及鉴定小鼠神经干细胞 ,为相关的实验研究奠定基础。方法 利用神经干细胞条件培养基和单细胞克隆技术 ,从胚胎小鼠大脑皮质分离并体外培养胚胎期小鼠大脑皮质细胞 ,连续稳定传代 2 0代后 ,以免疫荧光细胞化学法检测克隆细胞Nestin和分化后细胞中胶质原纤维酸性蛋白、β tublin和半乳糖苷酶的表达。 结果 从小鼠大脑皮层分离的细胞群具有连续形成克隆能力 ,其Nestin表达阳性。分化后的细胞可表达神经元、星形胶质细胞和寡突胶质细胞的特异性抗原。结论 成功分离并获得了小鼠皮层神经干细胞 ,该细胞可连续稳定传代 ,具备多向分化潜能 。
Objective To isolate, cultivate and identify the neural stem cells from the cortex of embryo mouse in vitro for the further related research.Methods Cerebral hemisphere of mouse fetuses were taken out. Suspension of cerebral cells was made. Neural stem cells were identified by immunofluorescent cytochemistry to the primary culture. Neural stem cells were isolated and cultured in special culture medium using single cell clone culture technique. After the stem cells were stably cultured successively for 20 generations, nestin, glial fibrillary acidic protein, β tublin, and galactosidase (Galc) were detected by indirect immunofluorescence cytochemistry. Results The stem cells isolated from the cerebral hemisphere of mouse embryo with positive expression of nestin could be passaged in vitro for at least 20 generations. Inoculated on the coverglass with poly 1 ornithine and cultured in medium containing serum, the stem cells were induced to differentiate. The cells differentiated from these stem cells showed green flurescence by GFAP fluorescent staining, red fluorescence by β tublin fluorescent staining, or green fluorescence by Galc fluorescent staining. Conclusion The stem cells from cortex of mouse embryo can be successfully isolated and cultured in vitro. The stem cells can be passaged in series and possessed the potential of multi directional differentiation and express the specific antigens of neuron, astrocytes, and oligodendrocytes. Thus they can be used for further experimental study.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2002年第12期832-835,共4页
National Medical Journal of China
基金
国家自然科学基金资助项目 (3 9970 75 2 )
