Objective.To construct the eukaryotic expression vector hypoxia-inducible factor 1α-pcDNA3.1and to investigate its transfective efficiency into mesenchymal stem cells (MSCs) in vitro and the expression of HIF-1αgene in MSCs.Methods.mRNA of Wistar Rats' myocardial cells was extracted, and cDNA was synthesized with Reverse Transcription Kit, HIF-1αwas amplified by polymerase chain reaction (PCR), and constructed intopcDNA3.1. Transfected HIF-1α-pcDNA3.1into MSCs by liposome mediated method. The expression of HIF-1αin the cells was detected by Western Blot Analysis and ELISA.Results.Eukaryotic expression vector HIF-1α-pcDNA3.1was constructed successfully. Analyzed by flow cytometer, The MSCs' surfaces mark were CD44+, SH3(CD73)+, CD34−, CD45−and the CD44+ cells and SH3(CD73)+ cells were 94.7% and 97.3%, respectively, showing the high purity of the cultured MSCs. After inducing, the cultured MSCs can differentiate into osteoblasts and adipocytes successfully. In HIF-1αgene transfected MSCs, the expression of HIF-1αmRNA and HIF-1αprotein were both increased obviously.Conclusion.HIF-1αwas cloned successfully. HIF-1α-pcDNA3.1can be transfected into MSCs by liposome-mediated method effectively and which resulting stable expression of HIF-1αin transfected MSCs.
Stabilization of Hypoxia-Inducible Factor-1 Alpha Augments the Therapeutic Capacity of Bone Marrow-Derived Mesenchymal Stem Cells in Experimental Pneumonia
