Introduction: Oxygen tension is one of the most important stimuli in stem cell biology. In this study, we investigate the considerable influence of hypoxia on CX43 gene expression as one of the most important gap junction on the surface of mesenchymal stem cells.
Methods: Mesenchymal stem cells were isolated from C57BL/6 mouse bone marrow and cultured in DMEM medium under low oxygen tension (1% pO2). Hypoxic conditions were applied in time table as 4, 8, 16, 24 and 48 hours and reoygenation immediately was applied for 8 hours. Then, RNA samples were isolated and cDNAs were constractedfor gene expression quantification using Real-Time PCR technique. Finally, the data were analyzed by statistical methods.
Results: the comparative threshold cycle method (ΔΔCT, ratio=2 –ΔΔCT) and statistical analysis showed that connexin43 gene expression had been increased in 4, 16 and 48 hours after applying hypoxic (8.61, 1.00 and 2.86, respectively) and was decreased after 8 and 24 hours hypoxic conditions
(0.03and 0.14) in comparision with normoxic cells (21% pO2).As well as the rate at Re-oxygenation for periods of 4, 8, 16, 24, and 48 h, is 3.9 , 0.59, 3.16 ,1.31 and 4.27 respectively.
Conclusoin: the alteration in oxygen tension effects on the extent of CX43 gene expression. Hypoxia and Re-oxygenation conditions resulted in the up and down-regulation of CX43 gene expression, respectively.
Keywords: Connexin 43, gene expression profile, hypoxia, Mesenchymal Stem Cell
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