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Compositions and methods for differentiating stem cells into cell populations comprising beta-like cells

  • US 9,371,516 B2
  • Filed: 09/18/2015
  • Issued: 06/21/2016
  • Est. Priority Date: 09/19/2014
  • Status: Active Grant
First Claim
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1. A method of culturing human pluripotent stem cells to produce a cell population comprising pancreatic β

  • -like cells comprising;

    (a) culturing the stem cells on an extracellular matrix for 2-4 days in a chemically defined medium, basic fibroblast growth factor, Activin A, and BMP4;

    (b) culturing the cells on an extracellular matrix from step (a) for 2-4 days in the presence of chemically defined insulin, transferrin and selenium (ITS) medium, FGF7, and nicotinamide;

    (c) culturing the cells on an extracellular matrix from step (b) for 3-5 days in the presence of a chemically defined ITS medium;

    retinoic acid;

    Noggin; and

    nicotinamide; and

    (d) culturing the cells on an extracellular matrix from step (c) for 6-10 days in the presence of a B27 serum-free medium, IGF I, IGF II, FGF7, insulin, nicotinamide, exendin-4, an ALK5i II, and forskolin, thereby producing insulin+glucagon−

    cells that exhibit in vitro glucose stimulated insulin secretion.

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