Abstract 16362: Serum-Free Differentiation and Enrichment of Functional Human Vascular Smooth Muscle Cells From Embryonic Stem Cells

Background: Currently, surface markers enabling cell sorting of viable vascular smooth muscle cells (SMCs) from mixed cell populations or tissues for therapeutic or diagnostic purposes are not available. Here we define a method to enrich for functional human vascular SMCs derived from human embryonic stem cells (hESCs). Methods & Results: Embryoid bodies (EBs) subjected to serum-free cardiovascular-directed differentiation were characterized over 30 days for SMC-specific gene expression by qRT-PCR, immunohistochemistry (IHC), and flow cytometry. EBs were composed of endothelial cells, fibroblasts, cardiomyocytes, and SMCs. Flow cytometry of day 28 EBs reveals that the SMCs form a distinct sub-population, which can integrate with endothelial cells in an in vitro angiogenesis assay. To enrich for SMCs, day 28 EBs were dissociated and cultured as monolayers. Passaging monolayers in serum-free conditions resulted in the loss of cardiomyocyte, endothelial and fibroblast populations. qRT-PCR and IHC analyses sh...