Abstract 302: Generation of Patient-Specific Tissue-Engineered Blood Vessels From Nonintegrated Induced Pluripotent Stem Cells

Abstract

Cell resourcing remains one of the major problems in the development of patient-specific TEBVs. The iPSC derived cardiovascular cells provide a promising source for TEBVs and cell therapy. The goal of this study is to construct TEBVs using patient-specific iPSCs. Using a non-integrated episomal vector based reprogramming system, we generated human iPSCs from peripheral blood mononuclear cells (PBMCs) isolated from the blood harvested from aneurysm patients with bicuspid aortic valves and coronary artery bypass patients with normal aortas and aortic valves. These iPSCs showed the expression of pluripotent markers and could differentiate into 3 layers of cells in vitro. We directed the iPSCs differentiation into vascular smooth muscle cells (VSMCs) in vitro. We documented the expression of VSMC markers, including smooth muscle α-actin (α-SMA), calponin and SM22α by quantitative real-time-PCR, immunocytochemistry, as well as Western-blotting. Furthermore, we confirmed that the iPSC-derived VSMCs could contract in response to carbachol, a muscarinic agonist. The treatment of iPSC-derived VSMCs with IL-1β, an atherogenic cytokine, changed MMPs (matrix metallopeptidases) activity. Finally, we found that iPSC-derived VSMCs could survive well and secrete collagen when transferred and cultured on PLLA scaffolds in vitro, suggesting a great cell sourcing for tissue-engineered blood vessels. Our data suggests that by using non-integrated iPSCs, we could efficiently differentiate patient-specific iPSCs into functional VSMCs, which may provide great cell sourcing for patient-specific TEBVs.