Culture medium for epithelial stem cells and organoids comprising the stem cells
AssigneeKoninklijke Nederlandse Akademie van Wetenschappen
InventorsToshiro Sato, Johannes C. Clevers, Meritxell Huch Ortega
Life of this patent
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Abstract
The invention relates to a method for culturing epithelial stem cells, isolated tissue fragments comprising the epithelial stem cells, or adenoma cells, and culturing the cells or fragments in the presence of a Bone Morphogenetic Protein (BMP) inhibitor, a mitogenic growth factor, and a Wnt agonist when culturing epithelial stem cells and isolated tissue fragments. The invention further relates to a cell culture medium comprising a BMP inhibitor, a mitogenic growth factor, and a Wnt agonist, to the use of the culture medium, and to crypt-villus organoids, gastric organoids and pancreatic organoids that are formed in the culture medium.
Claims40
(canceled)
An in vitro method for obtaining a pancreatic epithelial organoid, the method comprising:
- incubating a pancreatic epithelial stem cell or an isolated pancreatic tissue fragment comprising a pancreatic epithelial stem cell with an extracellular matrix;
- and
- culturing the pancreatic epithelial stem cell or isolated pancreatic tissue fragment for a length of time sufficient to be able to observe formation of a pancreatic organoid, in an animal or human cell culture medium comprising:
- a Bone Morphogenetic Protein (BMP) inhibitor,
- a mitogenic growth factor, and
- a Wnt agonist, and
- obtaining from the culture medium a pancreatic epithelial organoid.
An in vitro method for obtaining a pancreatic organoid, the method comprising:
- culturing epithelial stem cells or isolated pancreatic tissue fragments comprising pancreatic epithelial stem cells in contact with an extracellular matrix for a length of time sufficient to be able to observe formation of a pancreatic organoid, in a medium comprising Noggin, EGF and Keratinocyte Growth Factor or Fibroblast Growth Factor, any one of R-spondin 1 through R-spondin 4 and/or Wnt-3a, and further comprising one or more supplements selected from, B27, N2, and N-Acetylcystein;
- and
- obtaining from the culture medium a pancreatic organoid.
10-18. (canceled)
An in vitro method for obtaining a pancreatic epithelial organoid, the method comprising:
- culturing pancreatic epithelial stem cells or isolated pancreatic tissue fragments comprising pancreatic epithelial stem cells in contact with an extracellular matrix for a length of time sufficient to be able to observe formation of a pancreatic organoid, in an animal or human cell culture medium comprising:
- a Bone Morphogenetic Protein (BMP) inhibitor;
- a Wnt agonist;
- and
- Epidermal Growth Factor (EGF);
- and
- obtaining from the culture medium a pancreatic epithelial organoid.
An in vitro method for obtaining a pancreatic adenoma organoid, the method comprising:
- incubating a pancreatic epithelial adenoma stem cell or an isolated pancreatic tissue fragment comprising a pancreatic epithelial adenoma stem cell with an extracellular matrix, and
- culturing the pancreatic epithelial adenoma stem cell or isolated pancreatic tissue fragment for a length of time sufficient to be able to observe formation of an organoid, in an animal or human cell culture medium comprising:
- a Bone Morphogenetic Protein (BMP) inhibitor, and
- a mitogenic growth factor, and
- obtaining from the culture medium a pancreatic adenoma organoid.
Dependencies are inferred from the claim text.