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

• Main Authors: Hanjaya-Putra Donny, Wanjare Maureen, Gerecht Sharon, Vo Elaine Tuong
• Format: Patent
• Language: English
• Published: 30.08.2016
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

This invention relates, e.g., to a method for differentiating mammalian (e.g., human) pluripotent stem cells (PSCs) into smooth muscle-like cells (SMLCs) in vitro, comprising a) plating a single-cell suspension of PSCs that are smaller than 50 μm at a seeding concentration of about 5×104 cells/cm2-about 1×105 cells/cm2 onto a suitable surface, and culturing the cells under conditions which prevent the PSCs from aggregating and induce differentiation of the PSCs into vasculogenic progenitor cells; b) harvesting the cultured cells of step a) and separating them into a single cell suspension of cells that are smaller than 50 μm; and c) plating the single cell suspension of step b) at a seeding concentration of about 1×104 cells/cm2-about 5×104 cells/cm2 on a suitable surface, and culturing the cells in a differentiation medium that is supplemented with platelet-derived growth factor BB (PDGF-BB) and transforming growth factor-beta 1 (TGF β1), for a sufficient period of time to allow the vasculogenic progenitor cells to mature into SMLCs. In one embodiment of the invention, the cells from step c) are further subjected to a shear force of at least 1 dyne/cm2 for a time period sufficient to enhance differentiation, maturation and/or functionality of the cells.