Concurrent generation of functional smooth muscle and endothelial cells via a vascular progenitor

Smooth muscle cells (SMCs) and endothelial cells (ECs) are typically derived separately, with low efficiencies, fromhumanpluripotent stemcells (hPSCs).The concurrent generation of these cell typesmight leadtopotentialapplications in regenerativemedicinetomodel,elucidate,andeventually treatvascular diseases. Here we report a robust two-step protocol that can be used to simultaneously generate large numbers of functional SMCs and ECs from a common proliferative vascular progenitor population via a two-dimensional culture system. We show here that coculturing hPSCs with OP9 cells in media supplemented with vascular endothelial growth factor, basic fibroblast growth factor, and bone morphogenetic protein 4 yields a higher percentage of CD31⁺CD34⁺ cells on day 8 of differentiation. Upon exposure to endothelial differentiationmedia andSMdifferentiationmedia, these vascular progenitors were able to differentiate andmature into functional endothelial cells and smoothmuscle cells, respectively. Furthermore, we were able to expand the intermediate population more than a billionfold to generate sufficientnumbers ofECsandSMCs inparallel forpotential therapeutic transplantations.