مهندسی سلول های بنیادی مزانشیم تولید کننده TGFb-3 برای کاربردهای بالینی

Title

Engineering of Transforming Growth Factor Beta 3-producing Mesenchymal Stem Cells for Tissue Engineering Applications

English Abstract

Transforming growth factor beta 3 (TGF-β3) is a regulatory cytokine that affects cellular differentiation, adhesion and formation of extracellular matrix. It has a major role in palate and lungs development and regulates skin wound healing and scar formation. Delivery of TGF-β3 to the site of injury in lungs, skin or an epithelial tissue can regulate the regenerative process and the behavior of the contributing cells. As mesenchymal stem cells (MSCs) have been shown to promote regenerative activities in the above tissues, it was hypothesized that engineering of TGF-β3-secreting MSCs can add a new regenerative modality for repair of the above tissues. Rat TGF-β3 cDNA was cloned in the plasmid vector of pcDNA3.1 using competent E. coli cells. The cloned product was sequenced and matched with the reference sequence in GenBank website. Mesenchymal stem cells were isolated from rat femur using flushing method and culture on a cell culture-treated plastic surface. In flow cytometry, they were positive for CD90 and CD106 and were capable of differentiation into osteoblasts, chondroblasts and adipocytes. Using Lipofectamine 2000 transfection reagent, Mesenchymal stem cells were transfected with the cloned cDNA and expression of TGF-β3 protein was confirmed by Western blotting. The transfected MSCs preserved their ability for the triple differentiation into osteoblasts, chondroblasts and adipocytes. In this study, we have successfully engineered transforming growth factor beta 3-secreting mesenchymal stem cells with preservation of their stemness characteristics. There are several implications for application of these cells in regenerative medicine.

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