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 1 alpha-pcDNA(3.1) and to investigate its transfective efficiency into mesenchymal stem cells (MSCs) in vitro and the expression of HIF-1 alpha gene in MSCs. Methods. mRNA of Wistar Rats' myocardial cells was extracted, and cDNA was synthesized with Reverse Transcription Kit, HIF-1 alpha was amplified by polymerase chain reaction (PCR), and constructed into pcDNA(3.1). Transfected HIF-1 alpha-pcDNA(3.1) into MSCs by liposome mediated method. The expression of HIF-1 alpha in the cells was detected by Western Blot Analysis and ELISA. Results. Eukaryotic expression vector HIF-1 alpha-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-1 alpha gene transfected MSCs, the expression of HIF-1 alpha mRNA and HIF-1 alpha protein were both increased obviously. Conclusion. HIF-1 alpha was cloned successfully. HIF-1 alpha-pcDNA(3.1) can be transfected into MSCs by liposome-mediated method effectively and which resulting stable expression of HIF-1 alpha in transfected MSCs. Copyright (C) 2009 Jinfu Yang et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.