Method for prodcing cardiomyocytes from induced pluripotent stem cells in an integrated process

A method for generating a population of cardiomyocytes from induced pluripotent stem cells (iPSCs) in an integrated process, the method comprising:

• seeding the iPSCs on a modified surface of a modified cell culture substrate, the modified cell culture substrate comprising:
• a patterned polydimethylsiloxane (PDMS) substrate with a stiffness of 667 kPa, the patterned PDMS substrate comprising an imprinted pattern of human fetus cardiomyocytes on a cell-contacting surface of the patterned PDMS substrate;
• a first coating comprising a plurality of polydopamine molecules chemically attached to the cell-contacting surface of the patterned PDMS substrate, wherein the plurality of polydopamine molecules have a coating concentration between 47 μg/cm2 and 53 μg/cm2;
• and
• a second coating covering a top surface of the first coating, the second coating comprising a plurality of Laminin 511 E8 Fragment (LME8) molecules chemically attached to the first coating, wherein the plurality of LME8 molecules have a coating concentration between 0.4 μg/cm2 and 0.6 μg/cm2;
• culturing the seeded iPSCs on the modified surface of the modified cell culture substrate in an animal component-free culture medium until reaching a confluency of at least 80%;
• and
• differentiating the cultured iPSCs to the population of cardiomyocytes on the modified surface of the modified cell culture substrate comprising:
• inducing differentiation of the cultured iPSCs to a population of mesoderm cells by culturing the cultured iPSCs in a first growth medium comprising a Wnt activator;
• inducing differentiation of the population of mesoderm cells to a population of cardiac mesoderm cells by culturing the population of mesoderm cells in a second growth medium comprising a Wnt inhibitor;
• and
• inducing differentiation of the population of cardiac mesoderm cells to the population of cardiomyocytes by culturing the population of cardiac mesoderm cells in a third growth medium comprising Insulin.