Culture media and conditions for in vitro expansion and/or maturation of hepatocytes
AssigneeSatellite Biosciences, Inc.
InventorsSunil Kumar MALLANNA, Maxwell P. VAN BUSKIRK, Soujanya S. KARANTH
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Abstract
Provided herein are completely defined two-dimensional culture conditions, including culture supplements and requirements for an extracellular matrix, facilitating the expansion of and long-term maintenance of primary human hepatocytes, as well as culture conditions and media for maturation of hepatocytes.
Claims142
A method for culturing primary human hepatocytes, the method comprising the step of culturing one or more hepatocyte in contact with an extracellular matrix (ECM) in the presence of an expansion medium comprising a basal medium for human cells to which is added:
- one or more Wnt signaling activator, one or more receptor tyrosine kinase ligand, and one or more epithelial phenotype stabilizing agent.
A method for maturing a population of hepatocytes, the method comprising the step of maturing an expanded hepatocyte population in the presence of a maturation medium comprising a basal medium for human cells to which is added one or more hepatocyte maturation supplements.
The method of any one of claims 82-99, wherein the one or more maturation supplements comprises an antibiotic, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), GLUTAMAX™, ITS, a Notch inhibitor, an EGFR inhibitor, Oncostatin M, an anti-oxidant, a glucocorticoid, a pregnane X receptor (PXR) activator, a bile acid, a cAMP or cAMP analog, cholesterol, a thyroid hormone, a serum replacement component, or any combination of the foregoing.
An expansion medium comprising a basal medium for human cells to which is added one or more Wnt signaling activator, one or more receptor tyrosine kinase ligand, and one or more epithelial phenotype stabilizing agent.
A maturation medium comprising a basal medium for human cells to which is added one or more hepatocyte maturation supplements.
Dependencies are inferred from the claim text.