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目的 :观察大鼠神经干细胞的体外生长特性。方法 :利用无血清培养基悬浮培养 ,观察神经干细胞的体外生长过程。并比较不同接种密度 ,不同传代时间对神经干细胞生长的影响。用免疫细胞化学技术检测神经干细胞巢蛋白 (nestin)的表达 ;用吖啶橙 (AO) /溴化乙啶 (EB)检测细胞活性 ;用DAPI荧光染料标记细胞。结果 :神经干细胞聚合成细胞团生长 ,细胞团成分复杂。细胞接种密度以 5× 1 0 5个细胞 /ml组生长较好 ;原代细胞生长 3～ 4d后传代为宜。传代后 4d时可获得大量nestin阳性细胞。DAPI标记率为 1 0 0 %。结论 :细胞团成分复杂。神经干细胞要高密度接种 ,及时传代。DAPI可作为有效的体外标记物
Objective: To observe the in vitro growth characteristics of neural stem cells in rats. Methods: The suspension culture of serum-free medium was used to observe the growth of neural stem cells in vitro. The effects of different seeding density and passage time on the growth of neural stem cells were compared. Immunocytochemistry was used to detect the expression of nestin in neural stem cells. Acridine orange (AO) / ethidium bromide (EB) was used to detect the cell viability. The cells were labeled with DAPI fluorescent dye. Results: Neural stem cells aggregated into clusters of cells with complicated cell clusters. Cell inoculation density of 5 × 105 cells / ml group grew well; primary cell growth after 3 ~ 4d passage is appropriate. A large number of nestin positive cells were obtained 4 days after passage. DAPI labeling rate was 100%. Conclusion: The cell mass is complex. Neural stem cells to high-density inoculation, timely passage. DAPI can be used as a valid in vitro marker