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Method of producing pancreatic hormone-producing cells

  • US 9,157,069 B2
  • Filed: 08/08/2011
  • Issued: 10/13/2015
  • Est. Priority Date: 08/09/2010
  • Status: Active Grant
First Claim
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1. A method of producing human pancreatic hormone-producing cells, comprising subjecting human pluripotent stem cells to the following steps (1)-(6):

  • (1) a step of cultivating human pluripotent stem cells in a medium containing (+)-(R)-trans-4-(1-aminoethyl)-N-(4-pyridyl)cyclohexanamide dihydrochloride,(2) a step of cultivating the cells obtained in the aforementioned step (1) in a medium containing a 6-[[2-[[4-(2,4-dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)-2-pyrimidinyl]amino]ethyl]amino]nicotinonitrile or (2′

    Z,3′

    E)-6-bromoindirubin-3′

    -oxime,(3) a step of cultivating the cells obtained in the aforementioned step (2) in a medium containing 6-[[2-[[4-(2,4-dichlorophenyl)-5-(4-methyl-1H-imidazol-2-yl)-2-pyrimidinyl]amino]ethyl]amino]nicotinonitrile and activin(4) a step of forming a cell mass from the cells obtained in the aforementioned (3), and cultivating the cell mass in a suspension state in a medium(5) a step of cultivating the cells obtained in the aforementioned step (4) in a medium containing dorsomorphin, retinoic acid, 4-[4-(1,3-benzodioxol-5-yl)-5-(2-pyridinyl)-1H-imidazol-2-yl]-benzamide or hydrate thereof and basic fibroblast growth factor,(6) a step of cultivating the cells obtained in the aforementioned step (5) to produce human pancreatic hormone-producing cells.

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