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US 2023/0399623 A1Published application
Artifical liver organoids and methods of their production
OwnerGareth J. Sullivan, Sean Harrison +1inventors
InventorsGareth J. Sullivan, Sean Harrison, Richard Siller
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Abstract
The present invention relates to in vitro methods for production of liver organoids.
Claims45
An in vitro method for production of liver organoids comprising:
- a. suspending vertebrate pluripotent stem cells in a liquid base medium at a density of from 0.5 to 5.0 ml medium per one million pluripotent stem cells;
- b. incubating the suspended pluripotent stem cells for 12 to 30 hours under agitation so that the pluripotent stem cells form aggregates;
- c. incubating the aggregates from step b. in the presence of a WNT agonist for 12 to 30 hours;
- d. incubating the aggregates from step c. in medium without WNT agonist for 12 to 30 hours;
- e. incubating the aggregates from step d. in the presence of DMSO for 4 to 9 days;
- and
- f. incubating the aggregates from step e. in the presence of a glucocorticoid and an agonist of the hepatocyte growth factor receptor (HGFR) for from 10 to 200 days.
4 more elements
The method according to 7, wherein the medium in step c. comprises from 2 to 6 μM CHIR99021.
An artificial liver organoid comprising hepatocytes, neurons, macrovascular endothelial cells (MVEC), liver sinusoidal endothelial cell (LSEC), hepatic stellate cells, Kupffer cells and cholangiocytes, wherein the artificial liver organoid comprises at least one lumen lined with CD31+ cells.
Dependencies are inferred from the claim text.