Liver organoids with intrahepatic sympathetic nerves and methods of use thereof
AssigneeChildren's Hospital Medical Center
InventorsTakanori TAKEBE, Rie OUCHI
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Abstract
Disclosed herein are methods of producing liver organoids with intrahepatic sympathetic neurons, or sympathetic nerves, from pluripotent stem cells. These intrahepatic sympathetic neurons, or sympathetic nerves, are involved in fatty liver disease pathogenesis, and the liver organoids produced from the methods provided herein may be used as a model system for fatty liver disease and pharmaceutical screening.
Claims36
A liver organoid, wherein the liver organoid comprises hepatic stellate cells, hepatocytes, and sympathetic neurons.
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A method of producing the liver organoid containing sympathetic neurons, the method comprising:
- a) activating an FGF signaling pathway and a Wnt signaling pathway, and optionally inhibiting a BMP signaling pathway, in definitive endoderm cells (DE), for a first period of time;
- b) activating an FGF signaling pathway, a Wnt signaling pathway, and a retinoic acid (RA) signaling pathway, and optionally inhibiting a BMP signaling pathway, in the cells of step a), for a second period of time, thereby differentiating the DE to posterior foregut cells;
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- c) embedding the posterior foregut cells in a basement membrane matrix, and optionally inhibiting a BMP signaling pathway in the embedded posterior foregut cells for a third period of time;
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- d) culturing the posterior foregut cells for a fourth period of time to differentiate the posterior foregut cells to liver organoids;
- wherein a BMP signaling pathway is inhibited in step b) and/or step c).
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A method of making a liver organoid having a reduced number of, or which is free of, sympathetic neurons, the method comprising inhibiting sympathetic neuron survival in a liver organoid comprising sympathetic neurons, wherein said inhibition of sympathetic neuron survival ablates the sympathetic neurons from the liver organoid, thereby making a liver organoid having a reduced number of, or which is free of, sympathetic neurons.
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A kit comprising means for performing the method according to claim 40.
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Dependencies are inferred from the claim text.