Optmization of condition in oxygen-glucose deprivation/re-oxygenation models of HT22 cells
10.3760/cma.j.issn.1671-8925.2016.01.011
- VernacularTitle:HT22细胞糖氧剥夺/复氧模型建立条件的优化研究
- Author:
Lixi ZHAO
1
;
Wenzhu WANG
;
Tong ZHANG
;
Hui CHEN
;
Lixu LIU
Author Information
1. 100068 北京,中国康复科学所,神经损伤与康复北京市重点实验室
- Keywords:
HT22 cell;
Oxygen-glucose deprivation/re-oxygenation model;
Cell doubling time;
Cell death;
Cell morphology
- From:
Chinese Journal of Neuromedicine
2016;15(1):55-59
- CountryChina
- Language:Chinese
-
Abstract:
Objective To optimize the HT22 cell oxygen-glucose deprivation/re-oxygenation models in hippocampal neurons of mice,discuss the basis of model establishment and test the related parameters.Methods HT22 cell growth curve was plotted by cell counting and cell doubling time was calculated,and then,the oxygen-glucose deprivation time and re-oxygenation time were determined by LDH assay.The CCK-8 assay,cell counting,cell morphology and flow cytometry were performed to further optimize the parameters of the models.Results The HT22 cell doubling time was 17 h 18 min.The oxygen-glucose deprivation time was 18 h and the re-oxygenation was 20 h.CCK-8 assay indicated that absorbance in cells of oxygen-glucose deprivation 18 h/re-oxygenation 20 h (OGD 18 h/R20 h) group was 0.334±0.138,which was significantly lower than that in the control group (0.756±0.026,P＜0.05);cells in the OGD18 b./R20 h group were shrank obviously.Flow cytometry indicated that the percentage of cells in terminal apoptosis or non-viable non-apoptotic cells was 24.56％±5.76％ and that of viable apoptotic cells was 2.63％±0.55％ in the OGD18 h/R20 h group,which were significantly higher than those in the control group,respectively (8.13％±0.87％ and 0.80％±0.10％,/P＜0.05).Conclusion The HT22 cell oxygen-glucose deprivation/re-oxygenation models are successfully established;both oxygen-glucose deprivation time and re-oxygenation time are related to the cell doubling time,and OGD18 h/R20 h can preferably help the establishment;.
