For the treatment of acute and chronic liver failure, there is a critical need for both temporary and more permanent modes of liver support. Past biological approaches have been unsuccessful due largely to the inability to keep liver tissue functional in vitro for long periods. The long-term objective of the proposed research is to investigate and establish the factors that are necessary to maintain functional hepatocytes in vitro and to use these hepatocytes as the key components of an artificial liver system. The intended approach towards preservation of hepatocyte function is aimed at reconstitution of the histological organization of the liver from its isolated components and thereby reconstructing normal in vivo intercellular anatomic relationships in a culture system. Our preliminary results have indicated that, by simply sandwiching hepatocytes in between two layers of collagenous matrix, both morphology and protein secretory function are maintained for at least one month.
The specific aims of our future studies are to: (1) characterize, in detail, the cell functions that have been preserved in this system (i.e. transcriptional rates, translational rates, enzymatic detoxification kinetics, gluconeogenic capacity); (2) study the effects of various perturbations on this culture system (i.e. changes in media composition, addition of other extracellular matrix components, addition of sinusoidal cells); (3) develop methodology for transplantation of these hepatocytes in the collagen sandwich configuration into a peritoneal cavity for temporary liver support in acute, reversible liver failure in animal models; and (4) design and construct a continuous culture perfusion system to evaluate the dynamic behavior of the cultured hepatocytes for use within an extracorporeal device in chronic long-term liver support in animal models. The principles developed in this work will provide a rational basis for the design of an artificial liver device for treatment of acute and chronic liver failure.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK041709-05
Application #
2141877
Study Section
Surgery and Bioengineering Study Section (SB)
Project Start
1989-08-15
Project End
1995-04-30
Budget Start
1993-08-01
Budget End
1995-04-30
Support Year
5
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199
Chan, Christina; Berthiaume, Francois; Lee, Kyongbum et al. (2003) Metabolic flux analysis of cultured hepatocytes exposed to plasma. Biotechnol Bioeng 81:33-49
Chan, Christina; Berthiaume, Francois; Lee, Kyongbum et al. (2003) Metabolic flux analysis of hepatocyte function in hormone- and amino acid-supplemented plasma. Metab Eng 5:1-15
Chan, Christina; Berthiaume, Francois; Washizu, Junji et al. (2002) Metabolic pre-conditioning of cultured cells in physiological levels of insulin: generating resistance to the lipid-accumulating effects of plasma in hepatocytes. Biotechnol Bioeng 78:753-60
Washizu, J; Berthiaume, F; Chan, C et al. (2000) Optimization of rat hepatocyte culture in citrated human plasma. J Surg Res 93:237-46
Moghe, P V; Berthiaume, F; Ezzell, R M et al. (1996) Culture matrix configuration and composition in the maintenance of hepatocyte polarity and function. Biomaterials 17:373-85
Stefanovich, P; Matthew, H W; Toner, M et al. (1996) Extracorporeal plasma perfusion of cultured hepatocytes: effect of intermittent perfusion on hepatocyte function and morphology. J Surg Res 66:57-63
Foy, B D; Rotem, A; Toner, M et al. (1994) A device to measure the oxygen uptake rate of attached cells: importance in bioartificial organ design. Cell Transplant 3:515-27
Bhatia, S N; Toner, M; Tompkins, R G et al. (1994) Selective adhesion of hepatocytes on patterned surfaces. Ann N Y Acad Sci 745:187-209
Ezzell, R M; Toner, M; Hendricks, K et al. (1993) Effect of collagen gel configuration on the cytoskeleton in cultured rat hepatocytes. Exp Cell Res 208:442-52
Ryan, C M; Carter, E A; Jenkins, R L et al. (1993) Isolation and long-term culture of human hepatocytes. Surgery 113:48-54

Showing the most recent 10 out of 16 publications