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Pluripotent stem cell culture on micro-carriers

  • US 9,969,972 B2
  • Filed: 02/25/2015
  • Issued: 05/15/2018
  • Est. Priority Date: 11/20/2008
  • Status: Active Grant
First Claim
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1. A method to differentiate human pluripotent stem cells to insulin-expressing cells on micro-carriers comprising the steps of:

  • a. attaching the human pluripotent stem cells to a volume of dextran micro-carriers;

    b. differentiating the human pluripotent stem cells on the dextran microcarriers to definitive endoderm cells using Activin A;

    c. differentiating the definitive endoderm cells on the dextran microcarriers into pancreatic endoderm cells;

    d. differentiating the pancreatic endoderm cells on the dextran microcarriers into pancreatic endocrine cells; and

    e. differentiating the pancreatic endocrine cells on the dextran microcarriers into insulin-expressing cells, wherein the step of attaching comprises culturing the human pluripotent stem cells in medium containing a Rho kinase inhibitor at a concentration of between about 0.25 μ

    M to about 10 μ

    M.

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