Journal of Cytology & Histology

Hypoxic Preconditioning Attenuated Simulated Ischemia/Reperfusion Injury in Cultured Bone Marrow Stromal Cells

Abstract

LiHua Shen, ErBing Wu, Lin Cao, Jin Chen, Cuiping Xia, Yu Long and Xinsheng Ding

Aim: Bone marrow stromal cells (BMSCs) are a promising candidate for cell therapy in ischemic stroke. However, the majority of BMSCs are readily lost after transplantation because of apoptosis due to ischemia-reperfusion (I/R) injury. In the present study, we aimed to evaluate the effects of hypoxia preconditioning (HP) on simulated I/R injury in cultured BMSCs.
Methods: Four generations of BMSCs were used as subjects. BMSCs were divided into 6 groups, including normal group, I/R group(I/R followed 0 h HP), 2 h-HP group(I/R followed 2 h HP ), 4 h-HP group(I/R followed 4 h HP), 6 h-HP group (I/R followed 6 h HP) and 8 h-HP group(I/R followed 8 h HP) 2 h-HP group(I/R followed 2 h HP ), 4 h-HP group(I/R followed 4 h HP), 6 h-HP group (I/R followed 6 h HP) and 8 h-HP group(I/R followed 8 h HP). BMSCs were subjected to HP by exposing the cells to hypoxia (2%O2). After 12 h of reoxygenation, BMSCs were suffered from 3 h ischemia (<0.5% O2 and serum deprivation) and subsequent reperfusion (I/R). Cell viability, hypoxia-inducible factor (HIF) 1-α, caspase-3 and apoptosis were tested using MTT, ELISA and immunofluorescence staining respectively.
Results: There were no changes in cell viability after 2 h to 8 h HP in cultured BMSCs as compared to the normal group. But HP upregulated the content of HIF-1α in BMSCs and protected the BMSCs from later I/R injury, as evidenced by increased cell viability and decreased Caspase-3 release and apoptosis, especially in 8 h-HP group.
Conclusion: HP attenuated the apoptosis induced by I/R injury via co-regulating the expression of HIF-1α and Caspase-3 in BMSCs, and thereby might play a beneficial effect on cell therapy