Immortalization of Human Hepatocytes by Temperature-Sensitive Sv40 Large-T Antigen

Smalley, Matthew J.; Leiper, Kenneth A.; Tootle, Rosemary; McCloskey, P; O’Hare, Michael J.; Hodgson, Hjf

doi:10.1290/1071-2690(2001)037<0166:iohhbt>2.0.co;2

Cited by 20 publications

(10 citation statements)

References 7 publications

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“…Transplantation of animal models with immortalized hepatocytes is based on the hypothesis that an ideal alternative to the transplantation of primary human hepatocytes would be the use of a clonal cell line that could be grown in culture and exhibit the characteristics of differentiated nontransformed hepatocytes following transplantation. Human and rat hepatocytes have been transduced with a retroviral vector expressing the immortalizing simian virus 40 large T antigen (SV40Tag) gene 40, 41. This approach allows hepatocyte expansion in vitro , but transplantation of immortalized cells cannot currently be envisaged in clinical settings.…”

Section: Immortalizationmentioning

confidence: 99%

Hepatocyte transplantation in animal models

et al. 2009

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More than 30 years after the first hepatocyte transplant to treat the Gunn rat, the animal model for Crigler-Najjar syndrome, there are still a number of impediments to hepatocyte transplantation. Numerous animal models are still used in work aimed at improving hepatocyte engraftment and/or long-term function. Although other cell sources, particularly hepatic and extrahepatic stem cells, are being explored, adult hepatocytes remain the cells of choice for the treatment of liver diseases by cell therapy. In recent years, diverse approaches have been developed in various animal models to enhance hepatocyte transduction and amplification in vitro and cell engraftment and functionality in vivo. They have led to significant progress in hepatocyte transplantation for the treatment of patients with metabolic diseases and for bridging patients with acute injury until their own livers regenerate. This review presents and considers the results of this work with a special emphasis on procedures that might be clinically applicable. Liver Transpl 15:7-14, 2009. © 2008 AASLD. Received May 15, 2008; accepted September 5, 2008.The liver was among the first organs considered for strategies based on the transplantation of isolated cells. The first method for isolating hepatocytes from the liver involved collagenase perfusion in laboratory animals; it was initiated by Berry and Friend in 1969 1 and developed by Seglen in 1976.2 The first hepatocyte transplant was performed to treat the Gunn rat, the animal model for Crigler-Najjar syndrome, which is congenitally unable to conjugate bilirubin and consequently exhibits lifelong hyperbilirubinemia. The transplant resulted in a decreased plasma bilirubin concentration.3 Later, isolated hepatocytes were transplanted into rats with liver failure induced by dimethylnitrosamine. 4,5 These experiments demonstrated that hepatocyte transplantation could potentially be used for the treatment of liver failure and innate defects of liver-based metabolism. More than 30 years later, these models are still used in work to improve hepatocyte engraftment and/or function.

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Section: Immortalizationmentioning

confidence: 99%

Hepatocyte transplantation in animal models

et al. 2009

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“…Nevertheless, a porcine hepatocyte-based BAL device improved the clinical outcome of patients with acute liver failure (ALF) [4]. Efforts have thus turned towards the immortalization of human primary hepatocytes as a source of unlimited supplies of differentiated cells [5][6][7][8]. Immortalized human hepatocytes (IHH) were first reported through the transduction of proliferating primary cells with a gammaretroviral vector expressing the simian virus 40 large tumor antigen SV40TAg.…”

Section: Introductionmentioning

confidence: 99%

Treatment of acetaminophen-induced acute liver failure in the mouse with conditionally immortalized human hepatocytes

Nguyen

Mai

Villiger

et al. 2005

Journal of Hepatology

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“…Several prior studies used viral oncogenes or the catalytic subunit of telomerase reverse transcriptase for immortalization (Pfeifer et al 1993;Kobayashi et al 2000;Ray et al 2000;Smalley et al 2001;Wege et al 2003;Nguyen et al 2005). By contrast, the adult mouse hepatocyte culture described here did not require immortalization and was not obtained from hepatoma tissue (Aden et al 1979;Knowles et al 1980).…”

Section: Resultsmentioning

confidence: 99%

“…Previously, various strategies were used to establish hepatocyte cell lines including deriving cells from a TGFα transgenic mouse (Wu et al 1994) and immortalizing hepatocytes from adult and fetal liver tissue by transfection with the viral oncogene SV40 T antigen or the catalytic subunit of telomerase (Harris 1987;Pfeifer et al 1993;Kobayashi et al 2000;Ray et al 2000;Smalley et al 2001;Wege et al 2003;Nguyen et al 2005). Hepatocyte cell lines have also been established using co-culture with rat liver epithelial cells (Roberts et al 1994) and a collagen gel sandwich culture system (Kono et al 1995).…”

Section: Introductionmentioning

confidence: 99%

The establishment and characterization of immortal hepatocyte cell lines from a mouse liver injury model

Risal

Cho

Sylvester

et al. 2011

In Vitro Cell.Dev.Biol.-Animal

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Hepatocytes are an important research tool used for numerous applications. However, a short life span and a limited capacity to replicate in vitro limit the usefulness of primary hepatocyte cultures. We have hypothesized that in vivo priming of hepatocyte could make them more susceptible to growth factors in the medium for continuous proliferation in vitro. Here, a novel approach used to establish hepatocyte cell lines that included hepatocyte priming in vivo prior to culture with a 3,5-diethoxycarbonyl-1,4-dihydrocollidine diet was attempted. The cell line grew in a monolayer while maintaining a granular cytoplasm and a round nucleus. Electron microscopy displayed hepatocyte-like features including mitochondria, glycogen granules, and the presence of bile canaliculi. This cell line expressed many mature hepatocyte-specific genes including albumin, alpha1-antitrypsin, glucose 6-phosphatase, and tyrosine aminotransferase. Functional characteristic of hepatocytes like the ability to store glycogen, lipid, and synthesis of urea is well demonstrated by this cell line. These cells demonstrated anchorage dependent growth properties in soft agar and did not form tumors after transplantation into nude mice. This cell line can be sustained in culture for more than 100 passages (>1.5 years) without undergoing noticeable morphological changes or transformation. This novel method resulted in the establishment of an immortal, non-transformed hepatocyte cell line with functional characteristics that may aid research of cell metabolism, toxicology, and hepatocyte transplantation.

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Immortalization of Human Hepatocytes by Temperature-Sensitive Sv40 Large-T Antigen

Cited by 20 publications

References 7 publications

Hepatocyte transplantation in animal models

Hepatocyte transplantation in animal models

Treatment of acetaminophen-induced acute liver failure in the mouse with conditionally immortalized human hepatocytes

The establishment and characterization of immortal hepatocyte cell lines from a mouse liver injury model

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