Transfection of recombinant adenovirus Ad-EGFP/HIF-1α to rat endothelial progenitor cells in vitro
AIM: To investigate the feasibility of transfection of recombinant adenovirus Ad-EGFP/HIF-1α to rat endothelial progenitor cells (EPCs). METHODS: The mononuclear cells were isolated from rat bone marrow using percoll density gradient centrifugation, then induced with VEGF, bFGF and EGF. The expression of cell markers was assessed by immunocytochemistry. AdEGFP/HIF-lα was amplified in HNK293 cells, and then transfected EPCs in vitro. Expression of enhanced green fluorescent protein (EGFP) in infected EPCs was observed by fluorescence microscope, cell growth activity was detected by MTT, transfection efficiency was detected by flow cytometry, and expression of HIF-lα in EPCs was verified by RT-PCR analysis. RESULTS: After EPCs were infected with recombinant adenovirus, green fluorescence was found and increased gradually with the time in EPCs. The survival rates of EPCs had no significant change after transfection, and the transfection rate increased gradually with the time.Hypoxia inducible factor let (HIF-1α) was detected in infected cells by RT-PCR. CONCLUSION: The recombinant adenovirus Ad-EGFP/HIF-1α can infect EPCs effectively, without significant effect on activity of EPCs.
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维普
