Derivation of contractile vascular smooth muscle cells from human pluripotent stem cells

This protocol describes a method to generate smooth muscle-like cells (SMLCs) from human embryonic stem cells and induced pluripotent stem (iPS) cells. The SMLCs are further cultured in low-serum medium for 18 days to generate contractile SMLCs (Con-SMLCs). Contractile cells mimic the native state of vascular smooth muscle cells (vSMCs) found in vessel walls. When SMLCs are cultured in high-serum medium in the presence of TGFb and PDFG-BB for 18 days, they differentiate into "synthetic" smooth muscle cells, termed Syn-SMLCs. Compared to Syn-SMLCs, Con-SMLCs have a reduced proliferation rate, increased contractile phenotype, numerous and active caveolae with enlarged endoplasmic reticulum and abundant stress fibers and bundles. When transplanted subcutaneously, Con-SMLCs encapsulated in Matrigel plugs migrated to newly grown vasculature, where they produced elastin to stabilize the blood vessels.