大鼠脑组织细胞培养的形态学观察

MORPHOLOGICAL OBSERVATION ON CULTURED NERVE CELLS OF RAT BRAIN IN VITRO

为观察原代培养 SD大鼠大脑组织 ,包括神经干细胞在内的各种细胞的形态 ,取新生大鼠 ,无菌操作下取出整个大脑 ,制备成单细胞悬液 ,接种培养 ,免疫细胞化学鉴定各类细胞的同时用光镜和扫描电镜对不同阶段和种类的细胞进行形态学观察。结果显示 :在体外培养出神经元、胶质细胞和神经干细胞 ;神经细胞在体外的发育时程跟体内基本一致 ,神经干细胞在体外 2 0 d左右分化成熟 ,获取了神经细胞和干细胞体外培养的光镜和扫描电镜形态。结果提示 ,成体神经细胞体外分离、培养的条件关键点在于尽可能贴近自然 ,由此可保证细胞培养的质量 ,获得可靠的神经干细胞、神经元和胶质细胞。为各类神经细胞提供了形态学资料。

To observe the morphological changes of different cells including neural stem cells following the culture of cerebral tissue derived from Sprague-Dawley (SD) rat. Cerebral tissues was taken out from SD rats 2-24 h after birth and then dissociated into single-cell suspension by mechanical method. The cell suspension was cultured in serum-containing culture medium. The morphological changes at different stages were observed by light microscope and scanning electron microscope, and then identified by immunocytochemical staining. The results showed that neurons,glial cells and neural stem cells were obtained by in vitro culture. The developmental process of neuronal cells in vitro was similar with that in vivo, but the neural stem cells became matured 20 d following culture. The neural stem cells, neurons and glial cells were identified as positive for nestin, MAP, CNP and GFAP by immunocytochemistry respectively. The experiment was successed in finding a facile method of dissociation and culture of cerebral tissue in vitro. The key point to dissociate and culture neuronal cells and to obtain satisfactory neurons,glial cells and neural stem cells was to keep the nature of culture conditions. the present study provided the morphological data of miscellaneous nerve cells.