Method for inducing ureteric bud-like tissue
AssigneeKyoto University
InventorsKenji Osafune, Shinichi Mae, Makoto Ryosaka
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Abstract
An object of the present application is to provide a method for inducing the differentiation from pluripotent stem cells, particularly iPS cells and ES cells, into ureteric bud cells. Another object of the present application is to provide a system for producing ureteric bud-like tissue from pluripotent stem cells through each stage of differentiations, i.e. anterior primitive streak cells, anterior intermediate mesoderm cells and Wolffian duct cells. Yet another object of the present application is to provide a method for maintaining ureteric bud-like organoids. Another object of the present application is to provide a method for expansion-culturing ureteric bud-like tip tissue.
Claims20
A method for inducing an early Wolffian duct cell comprising the following steps:
- (0) culturing a pluripotent stem cell in the presence of an activator of activin receptor kinase 4,7, a glycogen synthase kinase 3 beta (GSK3β) inhibitor and a bone morphogenetic protein-4 (BMP4) under a two-dimensional culture condition to obtain an anterior primitive streak cell,
- (1) culturing the anterior primitive streak cell in the presence of a fibroblast growth factor 8 (FGF8), a retinoic acid receptor agonist, a transforming growth factor beta (TGFβ) inhibitor, and a BMP inhibitor under a two-dimensional culture condition to obtain an anterior intermediate mesoderm cell culture, and
- (2) culturing the anterior intermediate mesoderm cell culture in the presence of a GSK3β inhibitor, a BMP inhibitor, a FGF and a glial cell line-derived neurotrophic factor under a two-dimensional culture condition.
Dependencies are inferred from the claim text.