Cardiomyocyte production

摘要

Methods and composition for the production of cardiomyocytes from differentiation of pluripotent stem cells are provided. For example, in certain aspects methods including differentiating pluripotent stem cells in a large volume of suspension culture in the presence of ROCK inhibitors are described. In further aspects, methods for differentiation of stem cells into cardiomyocytes that overcome variability between different stem cell clones and different batch of culture medium are provided.

主权项

1. A method for producing human cardiomyocytes comprising the steps of:
a) culturing a selected human pluripotent stem cell clone having been prepared from a single pluripotent stem cell, the human stem cell clone being either a human embryonic stem cell clone or a human induced pluripotent stem cell clone; b) individualizing the cultured human pluripotent stem cells of step (a); c) culturing the individualized human pluripotent stem cells obtained from step (b) under defined conditions in suspension culture in an aggregate promotion media comprising fibroblast growth factor 2 (FGF2), and a Rho-associated protein kinase (ROCK) inhibitor or a myosin II inhibitor, to form cell aggregates; d) culturing said cell aggregates in cardiac differentiation medium comprising FGF2, Activin, and bone morphogenetic protein 4 (BMP4) to form cardiomyocytes; and e) obtaining the cardiomyocytes.