Experimental Study of the Effects of Marrow Mesenchymal Stem Cells Transfected with Hypoxia-Inducible Factor-1α Gene

Objective. To construct the eukaryotic expression vector hypoxia-inducible factor 1α-pcDNA(3.1) and to investigate its transfective efficiency into mesenchymal stem cells (MSCs) in vitro and the expression of HIF-1α gene in MSCs. Methods. mRNA of Wistar Rats' myocardial cells was extracted, and cDNA was synthesized with Reverse Transcription Kit, HIF-1α was amplified by polymerase chain reaction (PCR), and constructed into pcDNA(3.1). Transfected HIF-1α-pcDNA(3.1) into MSCs by liposome mediated method. The expression of HIF-1α in the cells was detected by Western Blot Analysis and ELISA. Results. Eukaryotic expression vector HIF-1α-pcDNA(3.1) was 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-1α gene transfected MSCs, the expression of HIF-1α mRNA and HIF-1α protein were both increased obviously. Conclusion. HIF-1α was cloned successfully. HIF-1α-pcDNA(3.1) can be transfected into MSCs by liposome-mediated method effectively and which resulting stable expression of HIF-1α in transfected MSCs.