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Compositions comprising human embryonic stem cells and their derivatives, methods of use, and methods of preparation

  • US 9,488,643 B2
  • Filed: 10/25/2013
  • Issued: 11/08/2016
  • Est. Priority Date: 03/07/2006
  • Status: Active Grant
First Claim
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1. A method for testing the effect of a compound on hES cells, derivatives of hES cells, or combinations thereof, wherein the derivatives of hES cells are selected from the group consisting of hematopoietic stem cell progenitors, neuronal stem cell progenitors, mesenchymal stem cell progenitors, insulin producing stem cell progenitors, hepatocytic stem cell progenitors, cardiac stem cell progenitors, epithelial stem cell progenitors, and combinations thereof, and wherein the hES cells and derivatives of hES cells are obtained by culturing in media that is free from feeder cells, animal products, growth factors, leukemia inhibiting factor, fibroblast growth factor, vitamin supplements, membrane associated steel factor, soluble steel factor, and conditioned media, and further by method (1),wherein method (1) comprises expanding the cells free of animal products, feeder cells, growth factors, leukaemia inhibitory factor, supplementary mineral combinations, amino acid supplements, vitamin supplements, fibroblast growth factor, membrane associated steel factor, soluble steel factor, and conditioned media, comprising:

  • (i) introducing the cells in a cell culture medium consisting of minimal essential medium, a progestin, and a β

    -human chorionic gonadotropin (β

    hCG) agonist; and

    (ii) incubating the cells at a temperature of about 34°

    C. to about 38°

    C. in an environment of about 3.5% to about 6% carbon dioxide for about 12 hours to about 48 hours; and

    in the presence of the compound and determining the effect of the compound on the cells.

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