Construction of a differentiated human hepatocyte cell line expressing the herpes simplex virus-thymidine kinase gene.
Transient support using a hybrid artificial liver (HAL) device is a promising treatment for the patients with acute liver failure. Primary human hepatocytes are an ideal source for HAL therapy; however, the number of human livers available for hepatocyte isolation is limited by competition for use in whole organ transplantation. To overcome this problem, we previously established a highly differentiated human fetal hepatocyte cell line OUMS-29. Considering the potential risk when using these genetically engineered cells in humans, additional safeguards should be added to make the cells more clinically useful. In this work, the herpes simplex virus thymidine kinase (HSVtk) gene was retrovirally introduced into OUMS-29 cells. One of the HSVtk-expressed clones, OUMS-29/thymidine kinase (TK), grew in chemically defined serum free medium and expressed the genes of albumin, asialoglycoprotein receptor, glutamine synthetase, glutathione-S-transferase pi, and blood coagulation factor X. In vitro sensitivity of the cells to ganciclovir was evaluated. Intrasplenic transplantation of 50 x 10(6) OUMS-29/TK cells prolonged the survival of 90% hepatectomized rats compared with medium injection alone (control). In the present study, we have established highly differentiated immortalized human hepatocytes with tight regulation. The cells may be clinically useful for HAL treatment.
Kobayashi
N
N
First Department of Surgery, Okayama University Medical School, Japan.
Miyazaki
M
M
Westerman
K A
KA
Noguchi
H
H
Sakaguchi
M
M
Totsugawa
T
T
Watanabe
T
T
Matsumura
T
T
Fujiwara
T
T
Leboulch
P
P
Tanaka
N
N
Namba
M
M
D000418:Albumins / Q000096:biosynthesis; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004351:Drug Resistance / Q000235:genetics; D015774:Ganciclovir / Q000494:pharmacology; D015870:Gene Expression; D005814:Genes, Viral; D022781:Hepatocytes / Q000166:cytology* / Q000201:enzymology* / Q000637:transplantation / Q000821:virology; D006801:Humans; D017114:Liver Failure, Acute / Q000628:therapy; D019164:Liver, Artificial; D008297:Male; D051379:Mice; D008819:Mice, Nude; D051381:Rats; D011917:Rats, Inbred Lew; D018139:Simplexvirus / Q000201:enzymology* / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics*; D014161:Transduction, Genetic
D000418:Albumins; D013937:Thymidine Kinase; D015774:Ganciclovir