Differentiation of the Human Marrow Mesenchymal Stem Cells into Vascular Endothelium-like Cells in vitro
Aim: The two main obstacles of successfully grafting tissue engineered skin are the shortage of the resource of the seed cell and the long process of vascularization of grafted skin. This article investigates the feasibility and condition of differentiati
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Burn center, The first affiliated hospital of Nanchang University. Nanchang 330006, Jiangxi Province, P.R.China 2 First people's hospital of Zhengzhou city. Zhengzhou 450004, Henan Province, P.R.China
Abstract — Aim: The two main obstacles of successfully grafting tissue engineered skin are the shortage of the resource of the seed cell and the long process of vascularization of grafted skin. This article investigates the feasibility and condition of differentiation of the human marrow mesenchymal stem cells (MSCs) into the vascular endothelial-like cells in vitro. Methods: The MSCs were harvested , isolated and purified by combining density gradient centrifugation with adhering method. The cultured cells in induce group were treated with 10ȝg/L vascular endothelial growth factor(VEGF) and 5ȝg/L basic fibroblast growth factor(bFGF). No VEGF and bFGF were added in control group. The morphological changes of the cells were observed with inverted phase contrast microscope. The expression of CD34, Ϊ factor were determined by the immunocytochemistry methods. The transmission electron microscope was used to further explore the ultrastructure characteristics of the cultered cells. Results: The cells induced by VEGF and bFGF showed the morphological changes from long fusiform-shaped to short fusiform or flat shape, which was consistent with the significant characteristic of endothelium cells. The expression of CD34, Ϊ factor were positive in the induced cells, which were negative in the control group. The Weibel-Palade body could be found in the cytoplasm of some induced cells. Conclusion: Combined VEGF with bFGF can induce MSCs differentiate into vascular endothelial-like cells in vitro, which may be unfold the potential applied prospects for vascularization and application of tissue engineered skin. Keywords — mesenchymal stem cells, vascular endotheliallike cells, vascular endothelial growth factor, basic fibroblast growth factor.
I. INTRODUCTION Bone marrow mesenchymal stem cells (MSCs) are pluripotent stem cells present in bone marrow mesenchymal tissues. These cells have the capacity to differentiate into cells of multiple mesoderm-type lineages, including bone, fat, cartilage and muscle. MSCs seem to be the ideal candidates for tissue engineering , as they can be rapidly amplified and maintain pluripotent differentiation capabilities. The aim of our study was to test if MSCs were capable of being the new seed-cells for tissue engineering skin. In the present study, we have induced human MSCs to be differen-
tiated into vascular endothelium-like cells in vitro in the presence of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor( bFGF). This is significant in that it prepares us for future clinical application. II. MATERIALS AND METHODS A. Isolation and curture of bone marrow-derived human MSC The experiment was carried out in the institute of burn, the first affiliated hospital of Nanchang University. Bone marrow aspiration samples in this study were obtained sterilely from human donors w
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