目的:建立新生大鼠大脑皮层细胞原代培养体系并进行神经元前体细胞的增殖研究。方法:原代混合培养新生大鼠大脑皮层细胞;混合培养体系用血清培养基培养6d后换用无血清培养基培养,免疫细胞化学显色鉴定此培养过程中不同时间点的细胞类型,并存不同时间段加入BrdU,用于分析神经元前体细胞的增殖状况。结果:换用无血清培养基培养后,出现明显的细胞分层现象;神经元及其前体细胞的比例逐渐上升,而神经干细胞和星形胶质细胞的比例缓慢下降;增殖细胞的比例在O~24h、24~48h、48~72h时间段里逐渐增加,而在72~96h、96~120h时间段里逐渐减少。结论:新生大鼠大脑皮层细胞混合培养体系在无血清培养条件下促进神经元前体细胞的增殖。
Objective: To establish mixed primary culture system of cells from the cerebral cortex of newborn SD rats and study on the proliferation of neuronal precursor cells. Methods: Mixed primary cultures were prepared from the cerebral cortex of newborn SD rats; Prior to being cultured in serum-free medium, the mixed cells were maintained in serum medium for 6 days. Immunofluorescence staining was used to characterize the mixed cultures at different time points; BrdU was used to detect the proliferation of the neuronal precursor ceils. Results: After being cultured for 6 days, glial ceils were of the most abundant cell type in the primary mixed neuronal-glial cultures of newborn rat cortex and form a cell monolayer. After the serum medium was changed into the serum~free medium, neuronal precursor cells grew and proliferated on the top of the monolayer; The percentage of neurons and neuronal precursor increased significantly while that of neural stem cells and as- trocytes decreased~ The percentage of proliferative cells increased significantly in the first three days while it decreased in the following days. Conclusion: Mixed primary cultures in the serum-free medium can promote the proliferation of the neuronal precursor cells.