肾上腺嗜铬细胞瘤细胞(PC12 cells)去血清后,其凋亡与多巴胺能神经元的凋亡有许多共同之处,今通过研究神经干细胞(NSCs)对去血清诱导的PC12细胞凋亡的作用,进一步为NSCs移植治疗神经系统疾病提供相应的实验和理论依据。将正常培养的PC12细胞与NSCs以不同的方式进行去血清共培养,观察PC12细胞的形态,检测PC12细胞的活性,计算PC12细胞的存活率,同时检测培养基中胶质细胞源神经营养因子(GDNF)的浓度,分析不同培养方式下FNSCs对去血清诱导凋亡的PC12细胞的作用以及NSCs与去血清诱导凋亡的PC12细胞共培养后,其分泌GDNF的能力。结果表明:①去血清诱导PC12细胞凋亡呈时间依赖性,去血清72h后,PC12细胞存活的比率为44.25%:②NSCs培养基对去血清诱导凋亡的PC12细胞没有明显的保护作用;:③NSCs培养上清及NSCs对去血清诱导凋亡的PC12细胞具有保护作用;④NSCs与去血清诱导凋亡的PC12细胞共培养后,分泌GDNF的能力增强。
The apoptosis of pheochromocytoma cells line 12 (PC12 cells) induced by serum deprivation is similar to that of dopaminergic neuron cells. In this paper, the potential cytoprotective effects of neural stem cells (NSCs) on the apoptosis of PC12 cells induced by serum deprivation were investigated to provide experimental and theoretical basis for the therapies of nervous system diseases via NSCs transplantation. The normal PC12 cells were co-cultured with NSCs in DMEM/F12 medium free of serum and then PC12 cell morphologies, viabilities and survival were observed with inverted microscope and assessed with CCK-8 assay, respectively. Besides, the concentrations of glial derived neurotrophic factor (GDNF) in different media were detected with GDNF El isa kit. Finally, the effect of NSCs on PC12 cell apoptosis induced by serum deprivation and the NSCs' ability to secrete GDNF after co,culture were analyzed. The results show that:① PC12 cell apoptosis induced by serum deprivation increases with time and after 72 h, only about 44.25% PC12 cells survive; ② NSCs culture medium protects against PC 12 cell apoptosis insignificantly; ③ NSCs' supernatant and NSCs prevent PC12 cells from apoptosis mildly; ④ the amount of GDNF secreted by NSCs increases after co-culture the NSCs with the apoptotic PC 12 cells induced by serum deprivation.