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

摘要

Objective. To construct the eukaryotic expression vector hypoxia-inducible factor 1α-pcDNA3.1 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 pcDNA3.1. Transfected HIF-1α-pcDNA3.1 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-pcDNA3.1 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 were94.7% and 97.3%, respectively, showing the high purity ofthe cultured MSCs. After inducing, the cultured MSCs candifferentiate into osteoblasts and adipocytes successfully. InHIF-1α genetransfected MSCs, the expression ofHIF-1α mRNA andHIF-1α protein wereboth increased obviously. Conclusion.HIF-1α was clonedsuccessfully. HIF-1α-pcDNA3.1 can be transfected into MSCs by liposome-mediatedmethod effectively and which resulting stable expression ofHIF-1α in transfectedMSCs.