Smooth muscle-like cells (SMLCs) dervided from human pluripotent stem cells

A method for differentiating human embryonic stem cells (hESCs) into human smooth muscle-like cells (SMLCs) in vitro, comprising

• a) plating a single-cell suspension of hESCs that have been filtered through a 40 μm strainer, to generate a population of cells that are smaller than 40 μm, at a seeding concentration of about 5×104 cells/cm2, onto a collagen IV coated plate, and culturing the cells in a differentiation medium that comprises about 10% serum, for about 6 days,
• b) harvesting the cultured cells of step a) and filtering them through a 40 μm strainer to generate a single cell suspension of cells that are smaller than 40 μm;
• c) plating the single cell suspension of step b) at a seeding concentration of less than about 2×104 cells/cm′ on a collagen IV coated plate, and culturing the cells in a differentiation medium comprising about 10% (v/v) of serum and that is supplemented with about 10 ng/ml of PDGF-BB and about 1 ng/ml of TGF β1, for about 6 days;
• and
• d) stretching the cells from step c) by subjecting the cells to a flow-induced shear stress for at least 48 hours;
• wherein the flow-induced shear stress is exerted in a flow chamber, and
• wherein the human SMLCs are characterized by at least one of up regulated PDGFR-B, up regulated Calponin, up regulated SMA, up regulated Ang-1, and/or reduced expression of fibronectin, as compared to human aorta vascular smooth muscle cells.