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US 10,597,633 B2Granted
Culture method for organoids
AssigneeKoninklijke Nederlandse Akademie van Wetenschappen
InventorsMeritxell Huch Ortega, Johannes Carolus Clevers, Sylvia Fernandez Boj
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Abstract
The invention relates to improved culture methods for expanding epithelial stem cells and obtaining organoids, to culture media involved in said methods, and to uses of said organoids.
Claims52
A method for culturing epithelial stem cells, wherein said method comprises:
- culturing one or more epithelial stem cells in contact with an extracellular matrix in the presence of a first expansion medium, the first expansion medium comprising a basal medium for animal or human cells to which is added:
- one or more receptor tyrosine kinase ligands, a Wnt agonist that is a Lgr5 agonist, a TGF-beta inhibitor and a cAMP pathway activator,
- wherein the TGF-beta inhibitor is able to reduce the activity of the TGF-beta signaling pathway that acts via Smad2 and/or Smad3.
A method for differentiating epithelial stem cells, wherein said method comprises:
- culturing the cells in a differentiation medium comprising a basal medium for animal or human cells to which is added EGF, gastrin, FGF , DAPT, dexamethasone, HGF and a TGF-beta inhibitor, wherein the TGF-beta inhibitor is able to reduce the activity of the TGF-beta signaling pathway that acts via Smad2 and/or Smad3.
A method for culturing epithelial stem cells, wherein said method comprises:
- culturing one or more epithelial stem cells in contact with an extracellular matrix in the presence of an expansion medium, the expansion medium comprising a basal medium for animal or human cells to which is added:
- one or more receptor tyrosine kinase ligands, a Wnt agonist that is a Lgr5 agonist, a TGF-beta inhibitor, and a BMP pathway activator,
- wherein the TGF-beta inhibitor is able to reduce the activity of the TGF-beta signaling pathway that acts via Smad2 and/or Smad3.
Dependencies are inferred from the claim text.