Establishment of A Cell Line From A Woodchuck Hepatocellular Carcinoma

Unoura, Masashi; Kobayashi, Kenichi; Fukuoka, Kenichi; Matsushita, Fumiaki; Morimoto, Hideo; Oshima, Tohru; Kaneko, Shuichi; Hattori, Nobu; Murakami, Seishi; Yoshikawa, Hiroshi

doi:10.1002/hep.1840050608

Cited by 20 publications

(6 citation statements)

References 11 publications

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“…This report provides evidence that sequences homologous to HBV DNA were consistently present in cultured hepatocytes for several weeks and existed predominantly and perhaps exclusively in extrachromosomal forms. This observation differs from previous reports on established human and woodchuck hepatoma cell lines (Aden et al, 1979;Alexander et al, 1978;Unoura et al, 1985). In such cases, only integrated forms of HBV were found.…”

Section: Discussioncontrasting

confidence: 95%

Maintenance of Woodchuck Hepatitis Virus Activity in Woodchuck Hepatocyte Primary Culture

Thézé

Gripon

Fourel

et al. 1987

Journal of General Virology

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SUMMARYPrimary cultures of non-proliferating hepatocytes isolated by the two-step collagenase perfusion method from woodchuck naturally infected with hepatitis virus (WHV) were used to study WHV propagation in vitro. Hepatocytes carrying WHV DNA exhibited a very high level of survival and retained their morphological characteristics for 2 to 3 months. Over this time, they were found to produce virusspecific proteins and release viral particles with DNA polymerase activity into the medium. Using Southern blot analysis and a recombinant hepatitis B virus DNA plasmid probe, intracellular and extracellular viral DNA was consistently detected. Only extrachromosomal forms of WHV DNA were observed and no integration could be demonstrated in the DNA of the cells. The WHV DNA patterns were repeatedly identical with a characteristic smear starting from 3.3 kb associated with other smaller DNA fragments which presumably represented intermediate replicative forms of viral DNA. Furthermore, dot blot hybridization of the total RNA revealed the presence of WHV-specific transcripts in cells after 3 weeks of culture. All these results are compatible with the maintenance of active WHV replication in vitro although it was somewhat reduced after the first day of culture. This provides a mammalian model for hepadnavirus replication studies in stable primary hepatocyte cultures.

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Section: Discussioncontrasting

confidence: 95%

Maintenance of Woodchuck Hepatitis Virus Activity in Woodchuck Hepatocyte Primary Culture

Thézé

Gripon

Fourel

et al. 1987

Journal of General Virology

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“…1). Production of albumin and integration of WHV DNA into cellular DNA were proved (Unoura et al, 1985).…”

Section: Preparation Of Woodchuck Hepatocellular Carcinomaderived Celmentioning

confidence: 99%

“…In this report, we determined the karyotype of woodchuck somatic cells by G-banding, then analyzed and compared with it the chromosomal abnormalities of a woodchuck hepatocel-Mar carcinoma-derived cell line (WH257GElO) (Unoura et al, 1985).…”

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confidence: 99%

Analysis of the chromosomal abnormalities of a woodchuck hepatocellular carcinoma-derived cell line (WH257GE10).

文昭¹

1987

Acta hepatologica Japonica

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The normal woodchuck karyotype was determined by the G-banding technique, and the chromosomal abnormalities of a woodchuck hepatocellular carcinoma-derived cell line (WH257GE10) were analyzed and compared with it. The normal woodchuck karyotype was 38,XY in a male, and 38,XX in a female. Autosomes were classified into 2 groups (A and B). Group A consisted of 13 pairs (1-13) of metacentric or submetacentric chromosomes, and group B consisted of 5 pairs (14-18) of telocentric or sub-telocentric ones. The X chromosome was sub-metacentric, and the Y chromosome was telocentric. In WH257GE10, the modal number was 36. Deletion of one of the chromosome 5 pair, of the chromosome 13 pair, and of the Y Chromosome was observed. Two marker chromosomes (MI and M3 were found. These markers showed the same G-banded patterns except for a small chromosome fragment at the distal part of the long arm of MI. The chromosome aberrations were very stable through successive passages in vitro. These findings suggest that the chromosomal abnormalities found in WH257GE10 might be associated with the pathogenesis of woodchuck hepatocellular carcinoma.

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“…Unfortunately, of the above human HCC cell lines, metastatic potential was rarely mentioned or demonstrated.For animal HCC cell lines, the establishment of such cell lines in rat [31] , in woodchuck [32][33][34] , and in chicken [35] have been reported.…”

Section: Hepatocellular Carcinoma Cell Linementioning

confidence: 99%

Metastatic human hepatocellular carcinoma models in nude mice and cell line with metastatic potential

Tang¹,

Sun²,

Tian³

et al. 2001

WJG

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Metastatic human HCC model is needed for the studies on mechanism and intervention of metastatic recurrence. By using orthotopic implantation of histologically intact tissues of 30 surgical specimens, a patient-like metastatic model of human HCC in nude mice (LCI-D20) and a low metastatic model of human HCC in nude mice (LCI-D35) have been established. All mice with transplanted LCI-D20 tumors exhibited e x t r e m e l y h i g h m e t a s t a t i c a b i l i t y i n c l u d i n g spontaneous metastasis to liver, lungs, lymph nodes and peritoneal seeding. Remarkable difference was also found in expression of some of the invasiveness related genes and growth factors between the LCI-D20 and LCI-D35 tumors. PAI-1 increased gradually following tumor progression in LCI-D20 model, and correlated with tumor size and AFP level. Phasic expression of tissue intercellular adhesion molecule-1 in this model was also observed. Using corneal micropocket model, it was demonstrated that the vascular response induced by LCI-D20 tumor was stronger than that induced by LCI-D35 tumor. Similar report on metastatic human HCC model in nude mice and human HCC cell line with metastatic potential was rarely found in the literature. This LCI-D20 model has been widely used for the studies on intervention of metastasis, including anti-angiogenesis, antisense approach, metallopro teinase inhibitor, differentiation inducer, etc. It is concluded that the establishment of metastatic human HCC model in nude mice and human HCC cell line with metastatic potential will provide important models for the in vivo and in vitro study of HCC invasiveness, angiogenesis as well as intervention of HCC recurrence.Subject headings hepatocellular carcinoma; metastasis; metastatic model; nude mice; cell line; experimental intervention; angiogenesis

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Establishment of A Cell Line From A Woodchuck Hepatocellular Carcinoma

Cited by 20 publications

References 11 publications

Maintenance of Woodchuck Hepatitis Virus Activity in Woodchuck Hepatocyte Primary Culture

Maintenance of Woodchuck Hepatitis Virus Activity in Woodchuck Hepatocyte Primary Culture

Analysis of the chromosomal abnormalities of a woodchuck hepatocellular carcinoma-derived cell line (WH257GE10).

Metastatic human hepatocellular carcinoma models in nude mice and cell line with metastatic potential

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