Expansion of human hepatocyte populations by a retroviral gene transfer of simian virus 40 large T antigen.
A hybrid artificial liver (HAL) could be used to treat acute liver failure or to serve as a temporary support until orthotopic liver transplantation is available. Primary human hepatocytes are ideal as a source of hepatic function in a HAL device. However, the worldwide shortage of human livers available for hepatocyte isolation severely limits this form of therapy. A possible alternative is to use a tightly regulated cell line that can be economically grown in culture to have differentiated liver function. In this work, human hepatocytes were immortalized with a retroviral vector SSR#69 expressing the genes of simian virus 40 large T antigen and herpes simplex virus-thymidine kinase. One of the resulting clones, NKNT-3 , showed the gene expression of differentiated liver function and were sensitive to the antiviral agent ganciclovir. When transplanted into the spleen of rats subjected to 90% hepatectomy, NKNT-3 cells prolonged the survival of 90% hepatectomized rats. The cells provide the advantages of unlimited availability, sterility, uniformity, and freedom from pathogens. This work represents a potential novel strategy for resolving the organ shortage that currently limits the use of primary human hepatocytes to develop a HAL.
Kobayashi
N
N
First Department of Surgery, Okayama University Medical School, Japan.
Westerman
K A
KA
Taguchi
T
T
Sakaguchi
M
M
Fujiwara
T
T
Urata
H
H
Kishimoto
N
N
Hayashi
N
N
Nakaji
S
S
Murakami
T
T
Leboulch
P
P
Tanaka
N
N
D058186:Acute Kidney Injury / Q000628:therapy; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D002454:Cell Differentiation; D002455:Cell Division; D002460:Cell Line; D002482:Cellulose; D004351:Drug Resistance / Q000235:genetics; D015774:Ganciclovir / Q000494:pharmacology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005814:Genes, Viral; D005822:Genetic Vectors; D022781:Hepatocytes / Q000166:cytology* / Q000276:immunology* / Q000637:transplantation / Q000821:virology; D006801:Humans; D019164:Liver, Artificial; D051379:Mice; D016513:Mice, SCID; D008863:Microspheres; D012190:Retroviridae / Q000235:genetics; D014183:Transplantation, Heterologous
D000952:Antigens, Polyomavirus Transforming; D002482:Cellulose; D015774:Ganciclovir