Mohsen Khaki - Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran-Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran.
Ali Hatef Salmanian - National Institute for Genetic Engineering and Biotechnology, Tehran. Iran.
Hamid Abtahi - Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran.
Ali Ganji - Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran-Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran.
Ghasem Mosayebi - Molecular and Medicine Research Center, Arak University of Medical Sciences, Arak, Iran-Department of Microbiology and Immunology, School of Medicine, Arak University of Medical Sciences, Arak, Iran.

Background: Vascular endothelial growth factor-A (VEGF-A), an endothelial cell-specific mitogen produced by various cell types, plays important roles in cell differentiation and proliferation. In this study we investigated the effect of recombinant VEGF-A on differentiation of mesenchymal stem cells (MSCs) to endothelial cells (ECs). Methods: VEGF-A was expressed in E. coli BL۲۱ (DE۳) and BL۲۱ pLysS competent cells with the pET۳۲a expression vector. Recombinant VEGF-A protein expression was verified by SDS-PAGE and western blotting. Mesenchymal stem cell differentiation to ECs in the presence of VEGF-A was evaluated by flow cytometry and fluorescence microscopy. Results: Recombinant VEGF-A was produced in E. coli BL۲۱ (DE۳) cells at ۰.۸ mg/mL concentration. Expression of CD۳۱ and CD ۱۴۴ was significantly greater, while expression of CD۹۰, CD۷۳, and CD۴۴ was significantly less, in MSCs treated with our recombinant VEGF-A than in those treated with the commercial protein (p < ۰.۰۵). Conclusions: Recombinant VEGF-A expressed in a prokaryotic system can induce MSCs differentiation to ECs and can be used in research and likely therapeutic applications.

Cell differentiation, Endothelial cell, Mesenchymal stem cell, Vascular Endothelial Growth Factor A