Tolerance of rat bone marrow mesenchymal stem cells overexpressing human heme oxygenase 1 to ischemia/hypoxia injury / 中国组织工程研究

ABSTRACT

BACKGROUND:Under ischemia/hypoxia microenvironment,very low survival rate of transplanted bone marrow mesenchymal stem cells (BMSCs) in the host limits its efficacy in the treatment of acute myocardial infarction.OBJECTIVE:To explore the tolerance of human heme oxygenase 1 (hHO-1) gene modified rat BMSCs to ischemia/hypoxia injury.METHODS:The rat BMSCs were transfected with hHO-1 recombinant adenovirus.Western blot assay was used to determinate the optimal time of hHO-1 protein expression.hHO-1 modified rat BMSCs were cultured in hypoxia and serum-free conditions that simulated ischemia/hypoxia microenvironment in vivo.Cell counting kit-8 and trypan blue staining were performed to detect the survival rate of BMSCs at 12,24,48,72 hours after culture under the ischemia/hypoxia microenvironment.Flow cytometry was used to detect apoptosis in BMSCs at 24 hours after culture under the ischemia/hypoxia microenvironment.RESULTS AND CONCLUSION:The expression of hHO-1 protein was highest at 4 days after transfection.Under the ischemia/hypoxia microenvironment for 12,24,48,72 hours,the survival rates of transfected BMSCs were significantly higher as compared with the untransfected cells (P ＜ 0.05),shown by the cell counting kit-8 and trypan blue staining.In addition,the results from flow cytometry showed that there was a higher survival rate of transfected BMSCs than untransfected cells at 24 hours of culture under the ischemia/hypoxia microenvironment (P ＜ 0.05).To conclude,hHO-1 modified rat BMSCs have stronger tolerance to the ischemia/hvpoxia microenvironment.