Treatment of surgically induced acute liver failure by transplantation of conditionally immortalized hepatocytes.
The shortage of human livers available for hepatocyte isolation limits its clinical application. The availability of cloned, conditionally immortalized hepatocytes that could be grown in culture but would lose their transformed phenotype and provide metabolic support upon transplantation would greatly facilitate the treatment of acute liver failure. Toward this goal, we transduced isolated Lewis rat hepatocytes using a replication-defective recombinant retrovirus capable of transferring a gene encoding a thermolabile mutant simian virus 40 T antigen (SV40ts). The cloned, immortalized hepatocytes proliferate at 33 degrees C. At the nonpermissive temperatures (37-39 degrees C), they stop growing and exhibit characteristics of differentiated hepatocytes. These cells did not produce tumors when transplanted in mice with severe combined immunodeficiency disease or in syngeneic rats. To induce acute liver failure, Lewis rats were subjected to 90% hepatectomy (Hpx) and given 5% oral dextrose. All rats that did not undergo hepatocyte transplantation died within 96 hr. Fifty percent of rats that received intrasplenic injection of 10 x 10(6) primary Lewis rat hepatocytes (G2, n=6) or 10 x 10(6) SV40ts-conditionally immortalized (SV40ts-ci) hepatocytes (G3, n=8) 1 day before 90% hepatectomy survived, whereas 80% of rats that received an intraperitoneal injection of 200 x 10(6) primary Lewis rat hepatocytes (G4, n=10) or 200 x 10(6) SV40ts-ci hepatocytes (G5, n=10) on the day of hepatectomy survived. Survival after intraperitoneal injection of a cellular homogenate of 200 x 10(6) primary Lewis rat (G7, n=9) or SV40ts-ci hepatocytes (G8, n=10) on the day of Hpx was 33% and 40%, respectively, whereas survival after intraperitoneal injection of 200 x 10(6) Lewis rat bone marrow cells (G6, n=7) was 29%. Thus, transplanted, conditionally immortalized hepatocytes can be as effective as primary hepatocytes in supporting life during acute liver insufficiency. This work represents the first step in developing an hepatocyte cell line that would partially alleviate the organ-donor shortage and could be of potential clinical value.
Nakamura
J
J
Department of Surgery, University of Nebraska Medical Center, Omaha 68198, USA.
Okamoto
T
T
Schumacher
I K
IK
Tabei
I
I
Chowdhury
N R
NR
Chowdhury
J R
JR
Fox
I J
IJ
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000032:analysis; D017690:Cell Transplantation / Q000401:mortality; D006498:Hepatectomy / Q000401:mortality; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000276:immunology; D017114:Liver Failure, Acute / Q000209:etiology / Q000628:therapy*; D008297:Male; D051379:Mice; D010537:Peritoneum; D051381:Rats; D013154:Spleen; D015996:Survival Rate; D016042:Transplantation, Heterotopic
D000952:Antigens, Polyomavirus Transforming