Method of producing pancreatic hormone-producing cells

摘要

Disclosed is a production method of pancreatic hormone-producing cells in a form that mimicks the pancreatogenesis, the method comprising subjecting stem cells to the following steps:
(1) cultivating stem cells in a medium containing a Rho kinase inhibitor,(2) cultivating the cells obtained in (1) in a medium containing a GSK3 inhibitor,(3) cultivating the cells obtained in (2) in a medium containing GSK3 inhibitor and an activator of activin receptor-like kinase-4,7,(4) forming a cell mass from the cells obtained in (3), and cultivating the cell mass in a suspension state in a medium,(5) cultivating the cells obtained in (4) in a medium containing a retinoic acid receptor agonist, an inhibitor of AMP-activated protein kinase and/or activin receptor-like kinase-2,3,6, an inhibitor of activin receptor-like kinase-4,5,7 and a cell growth factor, and(6) cultivating the cells obtained in (5).

主权项

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.