Establishment of rat models of oxygen glucose deprivation/reoxygenation in adult neural stem cellsin vitro
10.3760/cma.j.issn.1671-8925.2011.12.012
- VernacularTitle:大鼠成年海马神经干细胞体外氧糖剥夺/复氧模型的建立
- Author:
Ng She TAN
1
;
Jian CHEN
;
Yang GUO
;
Rui-Qing CHEN
;
Can LI
;
Zhen-Zhou CHEN
Author Information
1. 南方医科大学珠江医院
- Keywords:
Oxygen glucose deprivation/reoxgenation;
Neural stem cell;
Apoptosis;
Model
- From:
Chinese Journal of Neuromedicine
2011;10(12):1238-1242
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish simple,stable and reliable rat models of oxygen glucose deprivation/reoxgenation(OGD/R)in adult neural stem cells(NSCs)in vitro.Methods The NSCs from adult Fisher344 rats were cultured in serum-free medium and identified using nestin and DAPI immunofluorescent double staining.These cells were washed with a Earle′s balanced salt solution without glucose for 2 times,then,incubated for different periods(2,4,6,8 and 10 h)in a trigas incubator with an atmosphere of 1％ O2,5％CO2 and 94％ N2,98％ humidity at 37 ° C.And then,these cells were removed from the anaerobic incubator,washed,and added DEME/F12 containing bFGF supplement.A normoxic-normoglycemic control group was employed.Morphological assessment of NSCs was performed by light microscopy after re-oxgenation for 24 h; CCK-8 colorimetric method was used to determine the survival and proliferation of NSCs,and flow cytometry was employed to detect the apoptosis of NSCs.Results After the setting of oxygen glucose deprivation for 2 h,the OD value and the survival rate in the OGD cells were increased as compared with those in control group without significant difference(P＞0.05).While the morphological damage of NSCs aggravated gradually and the OD value decreased in OGD cells following the prolongation of times; under the setting of oxygen glucose deprivation for 6 h,the OD value in OGD cells obviously decreased as compared with that in the control group(P＜0.05); under the setting of oxygen glucose deprivation for 6 h,the survival rate obviously decreased and the apoptosis rate significantly increased in OGD cells as compared with that in the control group(P＜0.05); under the setting of oxygen glucose deprivation for 6 h,the apoptosis rate of NSCs excessed to 50％.Conclusion By means oftrigas incubator,simple,stable and reliable models of OGD/R in NSCs in vitro can be successfully established.
