Insertion of a suicide gene into an immortalized human hepatocyte cell line

N. Kobayashi, H. Noguchi, T. Totsugawa, T. Watanabe, T. Matsumura, Toshiyoshi Fujiwara, M. Miyazaki, K. Fukaya, M. Namba, N. Tanaka

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

For developing a bioartificial liver (BAL) device, an attractive alternative to the primary human hepatocytes would be the use of highly differentiated immortalized human hepatocytes with a safeguard. To test the feasibility, the primary human hepatocytes were immortalized by a plasmid SV3neo encoding simian virus 40 large T antigen (SV40Tag) gene. A highly differentiated hepatocyte line OUMS-29 was established. A suicide gene of herpes simplex virus-thymidine kinase (HSV-TK) was retrovirally introduced into OUMS-29 cells as a safeguard for clinical application. One of the resulting HSV-TK-positive cell lines, OUMS-29/ tk, grew in chemically defined serum-free medium with the gene expression of differentiated liver functions. OUMS-29/tk cells were 100 times more sensitive to ganciclovir compared with unmodified OUMS-29 cells in in vitro experiments. We have established a tightly regulated immortalized human hepatocyte cell line. Essentially unlimited availability of OUMS-29/tk cells may be clinically useful for BAL therapy.

Original languageEnglish
Pages (from-to)373-376
Number of pages4
JournalCell Transplantation
Volume10
Issue number4-5
Publication statusPublished - 2001

Fingerprint

Suicide
Hepatocytes
Genes
Cells
Cell Line
Viruses
Liver
Artificial Liver
Thymidine Kinase
Simplexvirus
Ganciclovir
Simian virus 40
Viral Tumor Antigens
Serum-Free Culture Media
Antigens
Gene expression
Plasmids
Availability
Gene Expression
Equipment and Supplies

Keywords

  • Herpes simplex virus-thymidine kinase
  • Immoralized human hepatocytes
  • Suicide gene

ASJC Scopus subject areas

  • Cell Biology
  • Transplantation

Cite this

Kobayashi, N., Noguchi, H., Totsugawa, T., Watanabe, T., Matsumura, T., Fujiwara, T., ... Tanaka, N. (2001). Insertion of a suicide gene into an immortalized human hepatocyte cell line. Cell Transplantation, 10(4-5), 373-376.

Insertion of a suicide gene into an immortalized human hepatocyte cell line. / Kobayashi, N.; Noguchi, H.; Totsugawa, T.; Watanabe, T.; Matsumura, T.; Fujiwara, Toshiyoshi; Miyazaki, M.; Fukaya, K.; Namba, M.; Tanaka, N.

In: Cell Transplantation, Vol. 10, No. 4-5, 2001, p. 373-376.

Research output: Contribution to journalArticle

Kobayashi, N, Noguchi, H, Totsugawa, T, Watanabe, T, Matsumura, T, Fujiwara, T, Miyazaki, M, Fukaya, K, Namba, M & Tanaka, N 2001, 'Insertion of a suicide gene into an immortalized human hepatocyte cell line', Cell Transplantation, vol. 10, no. 4-5, pp. 373-376.
Kobayashi N, Noguchi H, Totsugawa T, Watanabe T, Matsumura T, Fujiwara T et al. Insertion of a suicide gene into an immortalized human hepatocyte cell line. Cell Transplantation. 2001;10(4-5):373-376.
Kobayashi, N. ; Noguchi, H. ; Totsugawa, T. ; Watanabe, T. ; Matsumura, T. ; Fujiwara, Toshiyoshi ; Miyazaki, M. ; Fukaya, K. ; Namba, M. ; Tanaka, N. / Insertion of a suicide gene into an immortalized human hepatocyte cell line. In: Cell Transplantation. 2001 ; Vol. 10, No. 4-5. pp. 373-376.
@article{ace9bd5f6758481482621d901d8eed65,
title = "Insertion of a suicide gene into an immortalized human hepatocyte cell line",
abstract = "For developing a bioartificial liver (BAL) device, an attractive alternative to the primary human hepatocytes would be the use of highly differentiated immortalized human hepatocytes with a safeguard. To test the feasibility, the primary human hepatocytes were immortalized by a plasmid SV3neo encoding simian virus 40 large T antigen (SV40Tag) gene. A highly differentiated hepatocyte line OUMS-29 was established. A suicide gene of herpes simplex virus-thymidine kinase (HSV-TK) was retrovirally introduced into OUMS-29 cells as a safeguard for clinical application. One of the resulting HSV-TK-positive cell lines, OUMS-29/ tk, grew in chemically defined serum-free medium with the gene expression of differentiated liver functions. OUMS-29/tk cells were 100 times more sensitive to ganciclovir compared with unmodified OUMS-29 cells in in vitro experiments. We have established a tightly regulated immortalized human hepatocyte cell line. Essentially unlimited availability of OUMS-29/tk cells may be clinically useful for BAL therapy.",
keywords = "Herpes simplex virus-thymidine kinase, Immoralized human hepatocytes, Suicide gene",
author = "N. Kobayashi and H. Noguchi and T. Totsugawa and T. Watanabe and T. Matsumura and Toshiyoshi Fujiwara and M. Miyazaki and K. Fukaya and M. Namba and N. Tanaka",
year = "2001",
language = "English",
volume = "10",
pages = "373--376",
journal = "Cell Transplantation",
issn = "0963-6897",
publisher = "Cognizant Communication Corporation",
number = "4-5",

}

TY - JOUR

T1 - Insertion of a suicide gene into an immortalized human hepatocyte cell line

AU - Kobayashi, N.

AU - Noguchi, H.

AU - Totsugawa, T.

AU - Watanabe, T.

AU - Matsumura, T.

AU - Fujiwara, Toshiyoshi

AU - Miyazaki, M.

AU - Fukaya, K.

AU - Namba, M.

AU - Tanaka, N.

PY - 2001

Y1 - 2001

N2 - For developing a bioartificial liver (BAL) device, an attractive alternative to the primary human hepatocytes would be the use of highly differentiated immortalized human hepatocytes with a safeguard. To test the feasibility, the primary human hepatocytes were immortalized by a plasmid SV3neo encoding simian virus 40 large T antigen (SV40Tag) gene. A highly differentiated hepatocyte line OUMS-29 was established. A suicide gene of herpes simplex virus-thymidine kinase (HSV-TK) was retrovirally introduced into OUMS-29 cells as a safeguard for clinical application. One of the resulting HSV-TK-positive cell lines, OUMS-29/ tk, grew in chemically defined serum-free medium with the gene expression of differentiated liver functions. OUMS-29/tk cells were 100 times more sensitive to ganciclovir compared with unmodified OUMS-29 cells in in vitro experiments. We have established a tightly regulated immortalized human hepatocyte cell line. Essentially unlimited availability of OUMS-29/tk cells may be clinically useful for BAL therapy.

AB - For developing a bioartificial liver (BAL) device, an attractive alternative to the primary human hepatocytes would be the use of highly differentiated immortalized human hepatocytes with a safeguard. To test the feasibility, the primary human hepatocytes were immortalized by a plasmid SV3neo encoding simian virus 40 large T antigen (SV40Tag) gene. A highly differentiated hepatocyte line OUMS-29 was established. A suicide gene of herpes simplex virus-thymidine kinase (HSV-TK) was retrovirally introduced into OUMS-29 cells as a safeguard for clinical application. One of the resulting HSV-TK-positive cell lines, OUMS-29/ tk, grew in chemically defined serum-free medium with the gene expression of differentiated liver functions. OUMS-29/tk cells were 100 times more sensitive to ganciclovir compared with unmodified OUMS-29 cells in in vitro experiments. We have established a tightly regulated immortalized human hepatocyte cell line. Essentially unlimited availability of OUMS-29/tk cells may be clinically useful for BAL therapy.

KW - Herpes simplex virus-thymidine kinase

KW - Immoralized human hepatocytes

KW - Suicide gene

UR - http://www.scopus.com/inward/record.url?scp=0034866055&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034866055&partnerID=8YFLogxK

M3 - Article

VL - 10

SP - 373

EP - 376

JO - Cell Transplantation

JF - Cell Transplantation

SN - 0963-6897

IS - 4-5

ER -