Selective growth of epithelial-like clear cells from adult rat liver by short-term exposure to glucocorticoids in primary culture

Miyazaki, Masahiro; Wahid, Syarifuddin; Miyano, Keiko; Sato, Jiro

doi:10.1007/bf01851189

Cited by 5 publications

(3 citation statements)

References 12 publications

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“…We have already observed that the primary-cultured liver cells from adult rat revealed increased alpha-fetoprotein (AFP) production 6 days post inoculation which corresponded to the appearance and proliferation of epithelial-like clear cells [16]. The epithelial-like clear cells showed G6Pase and TAT activities even though the levels of these enzyme activities were very low as compared with those in rat liver [18]. We have also reported that cloned epithelial cells derived from adult and newborn rat liver cell lines had low levels of G6Pase and TAT activities and produced small amounts of albumin and AFP [27].…”

Section: Discussionmentioning

confidence: 95%

“…Morphological and functional features of epithelial-like clear cells resemble those of the cloned rat liver epithelial cells mentioned above [18,27]. Therefore, it might be thought that the epithelial-like clear cells originate from the partially differentiated hepatocytic precursor ceils (stem cells) in the periportal areas in the liver lobules [7,18,27].…”

Section: Discussionmentioning

confidence: 96%

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Comparison of cytologic and biochemical properties between liver cells isolated from adult rats by trypsin perfusion and those isolated by collagenase perfusion

et al. 1984

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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 amino-transferase, 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 X 10(-5)M) and insulin (10 micrograms/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.

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 96%

Comparison of cytologic and biochemical properties between liver cells isolated from adult rats by trypsin perfusion and those isolated by collagenase perfusion

et al. 1984

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“…Liver tissue is comprised of cell populations whose proliferations display a wide variation in sensitivity to glucocorticoids (24). The reproducible increase in c-myc mRNA levels observed in liver after glucocorticoid administration may represent a large increase in c-myc mRNA levels in a subpopulation of cells.…”

Section: Dexamethasone Regulation Of C-myc Mrna Levels In Estrogen-wimentioning

confidence: 99%

Rapid Inhibition of c-mycGene Expression by a Glucocorticoid in the Avian Oviduct

Rories¹,

Lau²,

Fink³

et al. 1989

Molecular Endocrinology

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The glucocorticoid dexamethasone (DEX) causes a rapid, reversible reduction in c-myc mRNA level in the oviducts of estrogen-treated, immature chickens. The c-myc mRNA level begins to decrease by 5 min after injection of 0.5 mg DEX, reaches a minimum of 10% of the control value by 30 min, and returns to 30-40% of the control value by 4 h post injection. This rapid effect of DEX on the c-myc mRNA level occurs in both diethylstilbestrol-stimulated and diethylstilbestrol-withdrawn oviducts. The effect is dose dependent, with reduction of the c-myc mRNA measured with as little as 10 micrograms DEX injection (0.03 micrograms/g BW). The effect of the steroid is gene specific with H2B histone mRNA displaying a significantly reduced response. The effect is also tissue specific with liver displaying an increase of 170% of control values in c-myc mRNA level by 30 min after injection of 0.5 mg DEX. The reduction of avian oviduct c-myc mRNA levels by DEX may play a role in glucocorticoid inhibition of cell proliferation in this tissue. The rapidity of the steroid effects on c-myc expression makes it likely that the steroid-induced reduction of c-myc mRNA levels represents a direct primary action of the steroid-receptor complex on the c-myc gene expression.

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Effect of phenobarbital on adult rat liver cells treated with 3?-methyl-4-dimethylaminoazobenzene in primary culture

Miyazaki

Handa

Sato

1985

J Cancer Res Clin Oncol

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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.

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Selective growth of epithelial-like clear cells from adult rat liver by short-term exposure to glucocorticoids in primary culture

Cited by 5 publications

References 12 publications

Comparison of cytologic and biochemical properties between liver cells isolated from adult rats by trypsin perfusion and those isolated by collagenase perfusion

Comparison of cytologic and biochemical properties between liver cells isolated from adult rats by trypsin perfusion and those isolated by collagenase perfusion

Rapid Inhibition of c-mycGene Expression by a Glucocorticoid in the Avian Oviduct

Effect of phenobarbital on adult rat liver cells treated with 3?-methyl-4-dimethylaminoazobenzene in primary culture

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