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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of cancer research and clinical oncology 110 (1985), S. 191-195 
    ISSN: 1432-1335
    Keywords: Primary culture of adult rat liver cells ; 3′-methyl-4-dimethylaminoazobenzene ; Phenobarbital ; Chromosomal abnormality ; Gamma-glutamyltranspeptidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The effect of phenobarbital (PB) on liver cells treated with 3′-methyl-4-dimethylaminoazobenzene (3′-Me-DAB) was studied using primary cultures of normal adult rat liver cells. Following a 1-day attachment period, primary liver cell cultures were treated with 0.24 mM 3′-Me-DAB for 6 days, and then treated with or without PB at 0.75, 1.5 and 3 mM for 19 days. Similarly, control cultures were treated with 0.5% dimethylsulfoxide (DMSO), a solvent for 3′-Me-DAB, for 6 days, and then treated with or without PB in the same way. Each treatment was done on 8 cultures. Chromosome analysis and cytochemical assay for gamma-glutamyltranspeptidase (GGT) activity were carried out on the carcinogen-treated and control cultures between 1 and 2 months after initiation of primary culture. Chromosomal abnormalities were detected in 23 of 32 carcinogen-treated cultures and also in 2 of 28 control cultures tested. However, GGT positive cells were detected only in the carcinogen-treated cultures at a frequency of 22/32. Of the 23 carcinogen-treated cultures with chromosomal abnormalities, 18 contained GGT positive cells. These results show a good correlation between chromosomal abnormality and acquisition of GGT activity at culture dish level. Furthermore, in the carcinogen-treated cultures, PB treatment caused a dose-dependent increase in the number of GGT positive cultures and in the percentage of GGT positive cells in each culture, and also caused a dose-dependent increase in the number of cultures with chromosomal abnormalities.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1433-8580
    Keywords: Primary culture of adult rat liver cells ; 3′-Methyl-4-dimethylaminoazobenzene ; Proliferation induction of epithelial-like clear cells ; Gamma-glutamyl transpeptidase ; Chromosomal abnormality
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Primary mass cultures of isolated liver cells, which were prepared from normal adult rat by a collagenase-liver-perfusion technique, were treated with 3′-methyl-4-dimethylaminoazobenzene (3′-Me-DAB) to study the production of transformed liver cells. Enzymatically isolated liver cells had high sensitivity to 3′-Me-DAB in primary culture. By 2- or 6-day treatment, mature hepatocytes remarkably decreased in their numbers due to cytotoxic effect of 3-Me-DAB, but thereafter active proliferation of epithelial-like clear cells was observed. Six-day treatment induced epithelial-like clear cells with gross chromosomal abnormalities, although 2-day treatment failed to induce transformed cells. However, the transformed epithelial-like clear cells with chromosomal abnormalities were negative for gamma-glutamyl-transpeptidase (GGT).
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1433-8580
    Keywords: Primary culture of adult rat liver cells ; Trypsin liver perfusion ; Collagenase liver perfusion ; Liver cell population ; Liver-specific functions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Primary cultures of liver cells isolated from adult rats by trypsin and collagenase perfusion techniques were carried out to compare cytologic and biochemical properties between the differently prepared cells. Trypsin-dispersed cells consisted of comparatively smaller cells, whereas collagenase-dispersed cells consisted of larger cells. The cell attachment efficiency on culture day 1 was about twice as high in the liver cells prepared with collagenase than those prepared with trypsin. Mature hepatocytes isolated by collagenase perfusion could be maintained in the primary culture for a longer period than those isolated by trypsin perfusion. Epithelial-like clear cells started to grow much earlier in the primary culture of the trypsin-dispersed liver cells than in that of the collagenase-dispersed liver cells. Earlier proliferation of epithelial-like clear cells could not be induced by in vitro trypsinization of the collagenase-dispersed liver cells. Both kinds of enzymatically prepared liver cells showed albumin production and exhibited glucose 6-phosphatase (d-glucose-6-phosphate phosphohydrolase, EC 3.1.3.9, G6Pase) and tyrosine aminotransferase (l-tyrosine: 2-oxoglutarate aminotransferase, EC 2.6.1.5, TAT) activities for 1 week in the primary culture. Albumin production was higher in the liver cells prepared with collagenase than those prepared with trypsin, whereas G6Pase activity was almost the same between them. TAT activity up to culture day 2 was about 3-fold higher in the liver cells prepared with collagenase than in those prepared with trypsin. Combined supplementation of dexamethasone (1 × 10−5 M) and insulin (10 µg/ml) consistently improved the cell attachment efficiency and was very effective in the maintenance of mature hepatocytes in both types. Furthermore, these hormones enhanced the albumin production and TAT activity in both types.
    Type of Medium: Electronic Resource
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