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Journal of Biomedicine and Biotechnology
Volume 2009, Article ID 128627, 10 pages
http://dx.doi.org/10.1155/2009/128627
Research Article

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

1Department of Cardiothoracic Surgery, Second Xiangya Hospital, Central South University, Hunan, Changsha 410011, China
2Department of Cardiothoracic Surgery, Hunan Children's Hospital, Hunan, Changsha 410007, China
3Department of General Surgery, Human Provincial People's Hospital, Hunan, Changsha 410005, China

Received 11 February 2009; Revised 9 April 2009; Accepted 15 April 2009

Academic Editor: Omar Benzakour

Copyright © 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.

Abstract

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