The objective is the immobilization of hepatocytes for use in an artificial liver device. Hepatocytes will be isolated by established techniques using collagenase digestion and immobilized in a sodium alginate gel matrix. This alginate gel will provide mechanical protection of the immobilized cells. The principles and technology developed in this work will provide a rational basis for design of treatment modalities for acute and chronic liver failure. The viability and function of the hepatocytes will be examined in the short and long term. The viability will be assessed using vital dye assays with both trypan blue and fluorescence dyes. The function of the hepatocytes will be assessed by measuring the reaction kinetics for the mixed function oxidase system (cytochrome P450), the urea cycle, and conjugating enzymes (UDP-glucuronyltransferase and sulfate transferase). In acute liver failure animal models, the immobilized cells will be used both as an intraperitoneal transplant and in an extracorporeal circuit to provide short term liver support. In the last 9 years, the applicant has completed the 5 year General Surgery residency training program at the Massachusetts General Hospital and also received a masters and a doctoral degree in Chemical Engineering at the Massachusetts Institute of Technology. He will apply the education in engineering and his clinical training in Surgery to the development of techniques for artificial liver support. Though the applicant has a background in research, this research was in engineering and not in biochemistry, cell biology, and physiology of digestive diseases. The applicant will require further training and development in learning these basic science skills to become an independent investigator in liver research. The sponsor of the applicant will be Dr. Kurt Isselbacher from the Medical Service of the Massachusetts General Hospital. Dr. Isselbacher will provide the environment necessary to develop basic science skills in liver biochemistry. The Surgical Service of the Massachusetts General Hospital and the Department of Surgery, Harvard Medical School, will provide the academic commitment and laboratory space. Dr. Paul Russell from the Surgical Service of the Massachusetts General Hospital will collaborate on transplant and immunological aspects of the project.

Project Start
1987-01-01
Project End
1991-12-31
Budget Start
1988-01-01
Budget End
1988-12-31
Support Year
2
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199
Ryan, C M; Carter, E A; Jenkins, R L et al. (1993) Isolation and long-term culture of human hepatocytes. Surgery 113:48-54
Dunn, J C; Tompkins, R G; Yarmush, M L (1992) Hepatocytes in collagen sandwich: evidence for transcriptional and translational regulation. J Cell Biol 116:1043-53
Bader, A; Borel Rinkes, I H; Closs, E I et al. (1992) A stable long-term hepatocyte culture system for studies of physiologic processes: cytokine stimulation of the acute phase response in rat and human hepatocytes. Biotechnol Prog 8:219-25
Ryan, C M; Yarmush, M L; Tompkins, R G (1992) Separation and quantitation of polyethylene glycols 400 and 3350 from human urine by high-performance liquid chromatography. J Pharm Sci 81:350-2
Dunn, J C; Tompkins, R G; Yarmush, M L (1991) Long-term in vitro function of adult hepatocytes in a collagen sandwich configuration. Biotechnol Prog 7:237-45
Carter, E A; Hatz, R A; Yarmush, M L et al. (1990) Injury-induced inhibition of small intestinal protein and nucleic acid synthesis. Gastroenterology 98:1445-51
Tompkins, R G; Hilton, J F; Burke, J F et al. (1989) Increased survival after massive thermal injuries in adults: preliminary report using artificial skin. Crit Care Med 17:734-40
Dunn, J C; Yarmush, M L; Koebe, H G et al. (1989) Hepatocyte function and extracellular matrix geometry: long-term culture in a sandwich configuration. FASEB J 3:174-7
Tompkins, R G; Remensnyder, J P; Burke, J F et al. (1988) Significant reductions in mortality for children with burn injuries through the use of prompt eschar excision. Ann Surg 208:577-85
Carter, E A; Tompkins, R G; Yarmush, M L et al. (1988) Redistribution of blood flow after thermal injury and hemorrhagic shock. J Appl Physiol 65:1782-8