A new approach to develop a biohybrid artificial liver using a tightly regulated human hepatocyte cell line.

N. Kobayashi, H. Noguchi, T. Watanabe, T. Matsumura, T. Totsugawa, Toshiyoshi Fujiwara, Takehito Taguchi, H. Urata, N. Kishimoto, N. Hayashi, S. Nakaji, K. A. Westerman, P. Leboulch, T. Murakami, N. Tanaka

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Currently patients with liver failure have been treated with a various liver support systems including a whole liver perfusion, a non-biological artificial liver, and a biohybrid artificial liver. In a hepatocyte-based bioreactor, porcine hepatocytes or transformed human liver tumor cells have been utilized because of the ease of preparation. According to the clinical data reported as of now, satisfactory results have not been obtained from the use of currently available liver support devices. One of the problems is limited availability of primary human liver cells for developing live support systems because of the shortage of human liver. To resolve this issue, human hepatocytes were immortalized with a retroviral vector SSR#69 which contained the genes of simian virus 40 large T antigen (SV40Tag) and herpes simplex virus-thymidine kinase (HSV-TK). One of the immortal cell lines, NKNT-3, showed the gene expression of differentiated liver functions, grew steadily in chemically defined serum-free CS-C medium, and doubled in number in about 48 hours. Essentially unlimited availability of NKNT-3 cells supports their clinical use for liver support devices. To realize the high density culture of NKNT-3 cells in a bioartificial liver device, we have developed cellulose microspheres (CMS) which contain cell adhesive GRGDS (Gly-Arg-Gly-Asp-Ser) peptides. Within 24 hours after starting a stirring suspension culture, GRGDS-CMS efficiently immobilized NKNT-3 cells. An electron microscopic examination demonstrated that NKNT-3 cells attached on GRGDS-CMS had well-developed mitochondria, rough reticulums, and villous extensions. In this article, we review the history of extracorporeal liver support systems and describe an attractive strategy for developing a novel extracorporeal liver assist device using NKNT-3 cells and GRGDS-coated cellulose microspheres.

Original languageEnglish
Pages (from-to)229-235
Number of pages7
JournalHuman cell : official journal of Human Cell Research Society
Volume13
Issue number4
Publication statusPublished - Dec 2000

Fingerprint

Artificial Liver
glycyl-arginyl-glycyl-aspartyl-serine
Hepatocytes
Cell Line
Liver
Microspheres
Cellulose
Equipment and Supplies
Reticulum
Simian virus 40
Thymidine Kinase
Viral Tumor Antigens
Liver Failure
Bioreactors
Simplexvirus
Adhesives
Suspensions
Mitochondria
Swine
Perfusion

ASJC Scopus subject areas

  • Cell Biology
  • Cancer Research

Cite this

A new approach to develop a biohybrid artificial liver using a tightly regulated human hepatocyte cell line. / Kobayashi, N.; Noguchi, H.; Watanabe, T.; Matsumura, T.; Totsugawa, T.; Fujiwara, Toshiyoshi; Taguchi, Takehito; Urata, H.; Kishimoto, N.; Hayashi, N.; Nakaji, S.; Westerman, K. A.; Leboulch, P.; Murakami, T.; Tanaka, N.

In: Human cell : official journal of Human Cell Research Society, Vol. 13, No. 4, 12.2000, p. 229-235.

Research output: Contribution to journalArticle

Kobayashi, N, Noguchi, H, Watanabe, T, Matsumura, T, Totsugawa, T, Fujiwara, T, Taguchi, T, Urata, H, Kishimoto, N, Hayashi, N, Nakaji, S, Westerman, KA, Leboulch, P, Murakami, T & Tanaka, N 2000, 'A new approach to develop a biohybrid artificial liver using a tightly regulated human hepatocyte cell line.', Human cell : official journal of Human Cell Research Society, vol. 13, no. 4, pp. 229-235.
Kobayashi, N. ; Noguchi, H. ; Watanabe, T. ; Matsumura, T. ; Totsugawa, T. ; Fujiwara, Toshiyoshi ; Taguchi, Takehito ; Urata, H. ; Kishimoto, N. ; Hayashi, N. ; Nakaji, S. ; Westerman, K. A. ; Leboulch, P. ; Murakami, T. ; Tanaka, N. / A new approach to develop a biohybrid artificial liver using a tightly regulated human hepatocyte cell line. In: Human cell : official journal of Human Cell Research Society. 2000 ; Vol. 13, No. 4. pp. 229-235.
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