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Effect of Neural Stem Cells on Apoptosis of PC12 Cells Induced by Serum Deprivation

机译:神经干细胞对血清剥夺诱导PC12细胞凋亡的影响

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Neural stem cells (NSCs) have a bright application prospect to be used to treat neurodegenerative diseases due to their capacity to give rise to the appropriate cell types when they are grafted.At present,however,the function of NSCs after transplantation is not quite ensured,whether to replace the degenerative cells or to secrete nutrient factors.On the other hand,pheochromocytoma cell line 12 (PC12) cells have been widely used for investigating Parkinson's disease (PD) since their apoptosis is similar to that of dopaminergic neuron cells.Therefore,the possible cytoprotective effects of NSCs on the apoptosis of PC12 cells induced by serum deprivation were investigated in this paper.PC12 cells were cocultured with NSCs in DMEM/F12 medium free of serum,and their morphologies,viabilities,and survival were observed with an inverted microscope and assessed with a CCK-8 assay.In addition,the concentrations of glial derived neurotrophic factor (GDNF) in different medium were detected with a GDNF Elisa kit,and the mechanism of NSCs protective effect on PC12 cell apoptosis induced by serum deprivation was analyzed.The results showed that (1) PC12 cell apoptosis induced by serum deprivation increased with time,and only about 44.25% PC12 cells survived after 72 h;(2) NSCs culture medium protected against PC12 cell apoptosis insignificantly;(3) NSCs' supernatant and NSCs mildly prevented PC12 cells from apoptosis;(4) the amount of GDNF secreted by NSCs increased after the coculture with the apoptotic PC12 cells induced by serum deprivation.It can be concluded that there exists clear interaction between NSCs and apoptotic PC12 cells,and that GDNF secretion from NSCs is one of the important mechanisms to prevent the apoptosis of PC12 cells.
机译:神经干细胞(NSCs)移植后具有产生合适细胞类型的能力,因此在治疗神经退行性疾病方面具有广阔的应用前景。但是,目前尚不能完全保证神经干细胞在移植后的功能。一方面,嗜铬细胞瘤细胞系12(PC12)细胞由于其凋亡与多巴胺能神经元细胞相似,因此已被广泛用于帕金森氏病(PD)研究。本文研究了NSCs对血清剥夺诱导的PC12细胞凋亡的可能的细胞保护作用。将PC12细胞与NSCs在无血清的DMEM / F12培养基中共培养,观察其形态,存活力和存活率。倒置显微镜并用CCK-8分析评估。此外,用HPLC检测不同培养基中神经胶质来源的神经营养因子(GDNF)的浓度。分析了GDNF Elisa试剂盒以及NSCs对血清剥夺诱导的PC12细胞凋亡的保护作用机理。结果表明:(1)血清剥夺诱导的PC12细胞凋亡随时间增加,仅72.72岁后存活率约44.25%。 h;(2)NSCs培养基对PC12细胞凋亡的保护作用不明显;(3)NSCs的上清液和NSCs轻度抑制PC12细胞的凋亡;(4)与凋亡PC12细胞共培养后,NSCs分泌的GDNF数量增加。结论:NSCs与凋亡的PC12细胞之间存在明显的相互作用,NSCs分泌的GDNF是阻止PC12细胞凋亡的重要机制之一。

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