The derivation and maturation of vascular smooth muscle cells from human pluripotent stem cells

Vascular diseases include conditions that cause artery, vein, or lymph vessel damage and disorders that affect blood circulation, such as cardiovascular disease, and peripheral vascular disease. A promising treatment for vascular diseases is cell therapy using human pluripotent stem cells (hPSCs) for vascular regeneration. The differentiation of hPSCs depends on biochemical cues (e.g. growth factors) as well as biomechanical cues (e.g. strain/stress). We first used the growth factors TGF‐β and PDGF‐BB to derive vascular smooth muscle cells (v‐SMCs) from hPSCs i.e. human embryonic stem cells and induced pluripotent stem cells. The application of uniaxial cyclic strain and serum starvation was utilized to induce maturation of the v‐SMC derivatives. v‐SMCs derivatives were analyzed using immunofluorescence staining for v‐SMC markers including SMA, calponin, SM22 and SM‐MHC. The production and secretion of Collagen 1 and Fibronectin was also investigated. Western blot and RT‐PCR quantified the proteins and mRNA levels of the various markers. . Our results indicate that short term culture and exposure to TGF‐β and PDGF‐BB promote a more contractile phenotype while long term exposure produces a more synthetic phenotype. Additionally, the application of strain has varying effects on the expression of different v‐SMC markers. This research was partially funded by the March of Dimes.