IN-VITRO VASCULARIZED TISSUE EQUIVALENTS FROM HUMAN EMBRYONIC STEM CELL-DERIVED ENDOTHELIAL AND VASCULAR SMOOTH MUSCLE CELLS
Vascularization is one of the key elements towards success in tissue engineering and cell therapy. One of the major obstacles challenging successful tissue engineering is the formation of functional vascular networks. Secondly, these applications require scalable production of homogeneous populations of vascular cells that include endothelial cells (ECs) and vascular smooth muscle cells (vSMCs). Human embryonic stem cells (hESCs) are virtually an unlimited source of differentiated cells including ECs and vSMCs. However, current differentiation strategies need improvisation in terms of efficiency, reproducibility, and use of xenogeneic (animal-derived) products. To obtain scalable amounts of clinically competent vascular cells, we aimed to develop differentiation protocols with minimal use of xenogeneic products. Further, we investigated the functionality of hESC-derived vascular cells to engineer in-vitro vascularized tissue equivalents (VTE). Under feeder-free and chemically-defined conditions, we demonstrate efficient differentiation of hESCs into vascular progenitor populations using sequential modulation of Wnt, FGF, BMP and VEGF signaling pathways. Further, we demonstrate the differentiation of CD34+CD31+ cells into homogeneous populations of arterial and venous ECs under serum-free conditions. Similarly, the CD34-CD31- are differentiated to vSMCs under smooth muscle differentiation conditions and characterized. A prerequisite to realize the full potential of these ECs and vSMCs is their ability to form blood vessels. We analyzed this potential by in-vitro 3D co-culture of the hESC-derived arterial ECs and vSMCs in a polyethylene-glycol-fibrin gel under serum-free conditions. Upon the 3D co-culture, the arterial ECs and vSMCs self-organize and mature into a microvascular network to form in-vitro VTE. In summary, we demonstrate the efficient differentiation of hESCs to ECs (arterial and venous) and vSMCs in relatively xenogeneic-free microenvironment; and 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
HUMAN EMBRYONIC STEM CELLS, ENDOTHELIAL CELLS, VASCULAR SMOOTH MUSCEL CELLS, TISSUE ENGINEERING, VASCULARIZED TISSUE EQUIVALENTS, ANGIOGENESIS
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