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Stem cell-derived hepatocytes in co-culture and uses thereof

  • US 10,266,806 B2
  • Filed: 01/14/2015
  • Issued: 04/23/2019
  • Est. Priority Date: 01/14/2014
  • Status: Active Grant
First Claim
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1. A composition comprising:

  • a population of human hepatocytes derived from induced human pluripotent stem cells and a population of 3T3-J2 fibroblasts in co-culture (population of hepatocytes in co-culture) in vitro;

    a culture substrate, wherein the cell populations are disposed in a micropattern on the culture substrate and the micropattern comprises a predetermined two-dimensional pattern of multiple microdots, the micropattern defined by a microdot diameter and a center-to-center spacing between each of any two neighboring microdots, wherein each microdot has a diameter of about 500 μ

    m and the center-to-center spacing between each of any two neighboring microdots is about 1200 μ

    m, and the microdots comprise the human hepatocytes derived from induced human pluripotent stem cells and the space between the microdots comprises the 3T3-J2 fibroblast population; and

    a layer of material comprising gelatinous protein mixture secreted by Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells disposed on the co-culture,wherein the population of hepatocytes in co-culture exhibits a higher level of cytochrome P450 3A4 (CYP3A4) enzyme activity by at least day 22 of culture as compared to a population of human hepatocytes derived from induced pluripotent stem cells and not co-cultured with a population of 3T3-J2 fibroblasts (population of hepatocytes not in co-culture).

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