Methods for producing enucleated erythroid cells derived from pluripotent stem cells

  • US 9,988,602 B2
  • Filed: 03/24/2016
  • Issued: 06/05/2018
  • Est. Priority Date: 05/06/2008
  • Status: Active Grant
First Claim
Patent Images

1. A method of producing human pluripotent stem cell-derived enucleated erythroid cells in vitro, comprising:

  • (a) providing a human pluripotent stem cell cultured in serum free media;

    (b) differentiating said pluripotent stem cell in a serum free media comprising bone morphogenic protein 4 (BMP-4) and vascular endothelial growth factor (VEGF), and producing embryoid bodies, wherein step (b) does not comprise culturing in a media comprising thrombopoietin (TPO), Flt3 ligand (FL), or stem cell factor (SCF);

    (c) culturing the embryoid bodies obtained from step (b) in a serum free media comprising at least basic fibroblast growth factor (bFGF) and methylcellulose, and producing a hemangioblast, non-engrafting hemangio cell, or blast cell; and

    (d) culturing said hemangioblast, non-engrafting hemangio cell, or blast cell in a serum free medium comprising at least erythropoietin (EPO), interleukin-3 (IL-3), SCF and methylcellulose;

    (e) co-culturing cells obtained by culturing under the conditions of at least step (d) with stromal cells or mesenchymal stem cells (MSCs) to produce human pluripotent stem cell-derived enucleated erythroid cells,wherein the human pluripotent stem cell-derived enucleated erythroid cells express β

    -globin.

View all claims
    ×
    ×

    Thank you for your feedback

    ×
    ×