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Method of in vitro differentiation of neural stem cells, motor neurons and dopamine neurons from primate embryonic stem cells

  • US 8,153,424 B2
  • Filed: 10/31/2007
  • Issued: 04/10/2012
  • Est. Priority Date: 10/03/2001
  • Status: Expired due to Fees
First Claim
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1. A method of creating a population of human ventral spinal progenitor cells, the method comprising the steps of:

  • (a) obtaining a synchronous population of cells that are cultured from human embryonic stem cells, that are characterized by an early rosette morphology and that are Pax6+/Sox1

    ;

    (b) culturing the cells of step (a) with a sufficient amount of retinoic acid in adherent culture, wherein the resulting cells are positive for Hoxb4 and are negative for at least one of Otx2 or Bf1;

    (c) expanding the resulting cells of step (b) in a suspension culture with a sufficient amount of retinoic acid; and

    (d) culturing the cells of step (c) in a suspension culture with a sufficient amount of retinoic acid and an agent selected from the group consisting of sonic hedgehog and purmorphamine, wherein at least 80% of the resulting cells are Olig2+, Nkx2.2+, Irx3+ and Pax7

    .

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