Method for producing dopaminergic neuron progenitor cell

A method for producing dopaminergic neuron progenitor cells comprising:

• (i) culturing human pluripotent stem cells (PSC) on a Laminin-511 E8 coated plate in a medium comprising a Sonic hedgehog (SHH) signal stimulating agent and bFGF for 6 to 48 hours to precondition and maintain the stem cells in an undifferentiated state,
• (ii) culturing the pluripotent stern cells obtained in step (i) on the Laminin-511 E8 in a medium containing a BMP inhibitor and a TGF-beta inhibitor for at least one day:
• (iii) replacing the medium from (ii) with a medium containing a BMP inhibitor, a TGF-beta inhibitor, a SHH signal-stimulating agent, and FGF8 and further culturing the cells for at least one day:
• (iv) replacing the medium from (iii) with a medium containing a BMP inhibitor, a TGF-beta inhibitor, a SHH signal-stimulating agent, FGF8, and a GSK3β inhibitor and further culturing the cells for at least one day;
• (v) replacing the medium from (iv) with a medium containing a BMP inhibitor and a GSK3β inhibitor and further culturing the cells for at least one day to produce a population of cells containing higher percentage of Corin- and/or leucine-rich repeats and transmembrane domains 1 (Lrtm1)-positive cells as compared to a population of Corin- and/or Lrtm1 positive cells produced from PSC that are not pre-conditioned in a medium comprising SHH signal-stimulating agent and bFGF;
• wherein steps (ii)-(v) are performed for 10-21 days;
• and
• (vi) collecting Corin positive and/or Lrtm1 positive cells and culturing the Corin positive and/or Lrtm1 positive cells in a suspension culture in a medium comprising one or more neurotrophic factors to produce dopaminergic neuron progenitor cells.