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US 10,472,610 B2Granted
Method for generating pancreatic bud cells and therapeutic agent for pancreatic disease containing pancreatic bud cells
AssigneeKyoto University
InventorsKenji Osafune, Taro Toyoda
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Abstract
Provided is a method for generating pancreatic bud cells, having the step of culturing PDX1+/NKX6.1− cells in a medium containing KGF, EGF and a BMP inhibitor. The culturing step may be performed in suspension cultures or in adherent cultures. When the cells are cultured in adherent cultures, the cells may be cultured in a medium further containing a ROCK inhibitor or a nonmuscle myosin II inhibitor.
Claims13
A method for generating pancreatic bud cells from pluripotent stem cells, comprising the following steps:
- (1) culturing the pluripotent stem cells in a medium containing an activin;
- (2) culturing the cells obtained in step (1) in a medium containing KGF;
- (3) culturing the cells obtained in step (2) in a medium containing KGF, a BMP inhibitor, a retinoic acid derivative and a hedgehog pathway inhibitor, wherein the steps (1)-(3) are under an adherent culture condition;
- and
- (4) dissociating the cells obtained in step (3) into single cells and culturing the dissociated cells under a suspension culture condition in a medium containing KGF, EGF and a BMP inhibitor.
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Dependencies are inferred from the claim text.