离体神经干细胞氧糖剥夺模型的建立及改变培养条件对神经干细胞的影响分析

摘要

ObjectiveTo establish a oxygen-glucose deprivation (OGD) model of neural stem cells (NSCs) and to investigate the effects of oxygen deprivation, glucose deprivation, OGD and high glucose on the survival and proliferation of NSCs culture respectively. Methods:The NSCs were separated from cortices of fetal mice and i-dentified. For the establishment of OGD, the NSCs were cultured in glucose-free DMEM and placed in a modular anaerobic chamber. The NSCs incubated in a normal condition were used as a control group. Both the cells in the control and OGD groups were incubated for different time durations (2 h, 4 h, 6 h, 8 h, 10 h). MTT assay was used to detect the effects of OGD on the survival and proliferation of NSCs. Besides, the NSCs were randomly as-signed into the control 8h group, oxygen deprivation 8 h group, glucose deprivation 8 h group, OGD 8 h group and high glucose 8 h group, and the cell viabilities were measured with MTT assay. Results:Compared to the nor-mal condition, the cell viabilities of OGD group were decreased at the time points of 8 h and 10 h significantly ( <0.05). The cell viabilities of oxygen deprivation 8h group, glucose deprivation 8h group and high glucose 8 h group were decreased when compared to control 8h group(all <0.05). There was no significant difference among glucose deprivation 8 h group, OGD 8 h group and high glucose 8 h group. Conclusion:The growth of NSCs was decreased significantly with OGD for 8 h or longer. In the model of OGD, low glucose concentrations might be the major detrimental factor for NSCs and the glucose concentration play an important role in the survival and proliferation of NSCs.%目的：建立离体神经干细胞（NSCs）氧糖剥夺（OGD）模型，并观察缺氧、缺糖等不同培养条件对NSCs存活和增殖的影响。方法：体外培养小鼠NSCs，制备OGD模型。将NSCs分为OGD组和正常组，每组分别干预2、4、6、8、10 h。采用MTT法检测OGD干预不同时间对NSCs活性的影响。通过MTT法观察正常培养、单独缺氧、单独缺糖、缺氧缺糖、高糖各组干预8 h对NSCs活性的影响。结果：OGD组干预8、10 h的NSCs OD值低于相应时点的正常组,差异有统计学意义（<0.05）。单独缺氧、单独缺糖、缺氧缺糖、高糖培养组的NSCs OD值均低于正常对照组（<0.05）。单独缺糖、高糖培养组与氧糖剥夺组的NSCs OD值比较，差异无统计学意义。结论：OGD≥8 h对NSCs造成明显的损伤。缺糖是OGD模型造成NSCs损伤的主要因素。葡萄糖浓度是影响NSCs存活和增殖的关键培养条件。