Long-term three dimensional tissue culture system
First Claim
1. A population of hepatocytes and nonparenchymal cells, derived using a method comprising:
- co-culturing hepatocytes and nonparenchymal cells, derived from disaggregated liver tissue, in the presence of (a) one or more growth factors that support the growth of hepatocytes comprising epidermal growth factor or hepatocyte growth factor and (b) beads coated with extracellular matrix protein that promotes cell adhesion under conditions sufficient to allow for the proliferation of said hepatocytes while retaining hepatic faction of said hepatocytes.
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Abstract
A method is provided for long term culture of proliferating hepatocytes that retain hepatic function to produce a hepatic cell culture. Hepatocytes and nonparenchymal cells are co-cultured ex vivo on a matrix coated with a molecule that promotes cell adhesion, proliferation or survival, in the presence of growth factors, resulting in a long-term culture of proliferating hepatocytes that retain hepatic function. The co-culturing method results in the formation of matrix/hepatic cell clusters that may be mixed with a second structured or scaffold matrix that provides a three-dimensional structural support to form structures analogous to liver tissue counterparts. The method can be used to form bio-articial livers through which a subjects blood is perfused. In an embodiment, the hepatocytes and nonparenchymal cells are derived from disaggregated liver tissue and are co-cultured in the presence of epidermal growth factor or heptocyte growth factor and beads coated with extracellular matrix protein, Alternatively, the hepatic cell culture may be implanted into the body of a recipient host having a hepatic disorder. Such hepatic disorders, include, for example, cirrhosis of the liver, induced hepatitis, chronic hepatitis, primary sclerosing cholangitis and alpha1 antitrypsin deficiency.
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Citations
6 Claims
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1. A population of hepatocytes and nonparenchymal cells, derived using a method comprising:
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co-culturing hepatocytes and nonparenchymal cells, derived from disaggregated liver tissue, in the presence of (a) one or more growth factors that support the growth of hepatocytes comprising epidermal growth factor or hepatocyte growth factor and (b) beads coated with extracellular matrix protein that promotes cell adhesion under conditions sufficient to allow for the proliferation of said hepatocytes while retaining hepatic faction of said hepatocytes.
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2. A method for generating a hepatic cell culture comprising:
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co-culturing hepatocytes and nonparenchymal cells derived from disaggregated liver tissue, in the presence of (a) one or more growth factors that support the growth of hepatocytes comprising epidermal growth factor or hepatocyte growth factor and (b) beads coated with extracellular matrix protein that promotes cell adhesion under conditions sufficient to allow for the proliferation of said hepatocytes while retaining hepatic function of said hepatocytes. - View Dependent Claims (3, 4, 5, 6)
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Specification