Establishment of Hypoxia /Reoxygenation Injury Model of Myocardial Cells in Rats by Using Anaerobic Incubator

Objective To explore a new way for establishment of a hypoxia/reoxygenation( H/R) injury model of myocardial cells line H9c2 in rats. Methods Myocardial cells line H9c2 were randomly divided into control group and experimental group. Control group was routinely cultured,while the experimental group was secondly divided into hypoxia for 2,4,8 and 12 h groups and posthypoxic reoxygenation for 2 h group. The cells in experimental group were cultured under the condition of 85% N2,10% H2,and 5% CO2with no-serous low carbohydrates of DMEM in hypoxia. The posthypoxic cells were entered into fresh DMEM and cultured in co2gas incubator for 2 h. The cell survival rate was detected by Trypan blue exclusion. The apoptotic rate was detected by flow cytometry. The content of lactate dehydrogenase( LDH) in the culture medium was detected by spectrophotometric method. Results Compared with those in control group,cell survival rate was significantly reduced,and the LDH content and apoptotic rate were significantly increased,and the differences were statistically significant( P 0. 05). Conclusion Establishment of H/ R injury model in rats by using anaerobic incubator is simple and practicable with good repeatability.