The present invention aims to provide a production methodof pancreatic hormone-producing cells in a form more mimickingthe pancreatogenesis.A method of producing pancreatic hormone-producing cells,comprising subjecting stem cells to the following steps (1) -(6):(1) a step of cultivating stem cells in a medium containing aRho kinase inhibitor(2) a step of cultivating the cells obtained in theaforementioned step (1) in a medium containing a GSK3inhibitor(3) a step of cultivating the cells obtained in theaforementioned step (2) in a medium containing GSK3 inhibitorand an activator of activin receptor-like kinase-4,7(4) a step of forming a cell mass from the cells obtained inthe aforementioned (3), and cultivating the cell mass in asuspension state in a medium(5) a step of cultivating the cells obtained in theaforementioned step (4) in a medium containing a retinoic acidreceptor agonist, an inhibitor of AMP-activated protein kinaseand/or activin receptor-like kinase-2,3,6, an inhibitor ofactivin receptor-like kinase-4,5,7 and a cell growth factor(6) a step of cultivating the cells obtained in theaforementioned step (5).
