Differential responsiveness of cultured suckling and adult rat hepatocytes to growth‐promoting factors: Entry into s phase and mitosis

Baribault, Hélène; Leroux-Nicollet, Isabelle; Marceau, Normand

doi:10.1002/jcp.1041220116

Cited by 31 publications

(16 citation statements)

References 34 publications

(53 reference statements)

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“…These conditions allow the majority of cells to undergo DNA synthesis and mitosis while still retaining measurable TAT activity and other hepatic functions (2). Hepatocytes and nonparenchymal cells in the primary cultures were discriminated by differential immunostaining for cytokeratin A and vimentin, respectively (1). In all cases, hepatocytes comprised >95% of the population.…”

Section: Methodsmentioning

confidence: 99%

Differential activity of a tissue-specific extinguisher locus in hepatic and nonhepatic cells.

Gourdeau¹,

Peterson²,

Fournier³

1989

Mol. Cell. Biol.

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Tissue-specific extinguisher 1 (Tse-1) is a genetic locus on mouse chromosome 11 that can repress expression of several liver genes in trans. This locus is clearly active in fibroblasts, as hepatoma cells retaining fibroblast chromosome 11 are extinguished for both tyrosine aminotransferase and phosphoenolpyruvate carboxykinase gene expression. To assess the activity of Tse-1 in other tissues, we transferred mouse chromosome 11 from several different cell types into rat hepatoma recipients. Tse-1 was active in nonhepatic cell lines derived from each primary germ layer, but Tse-1 activity was not apparent in hybrids between hepatoma cells and primary mouse hepatocytes. These differences in the genetic activity of murine Tse-1 were apparently heritable in cis.

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Section: Methodsmentioning

confidence: 99%

Differential activity of a tissue-specific extinguisher locus in hepatic and nonhepatic cells.

Gourdeau¹,

Peterson²,

Fournier³

1989

Mol. Cell. Biol.

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“…Hepatocytes cultured for various periods were treated with 0.12 mM colchicine for 8 h, fixed in methenokacetic acid (3:1), and then stained with Hoechst 33258 (Chen, 1977;Baribault et al, 1985). The proportion of hepatocytes in metaphase was evaluated by counting the percentage of hepatocyte mitoses in six separate fields (a total of about 500 cells).…”

Section: Mitosismentioning

confidence: 99%

“…EGF, insulin, and glucagon are growth-promoting factors of rat hepatocytes both in vivo (Bucher et al, 1978;Bucher and Weir, 1976;Bucher and McGowan, 1979) and in primary culture (Richman et al, 1976;McGowan et al, 1981;McGowan and Bucher, 1983;Friedman et al, 1981;Hasegawa et al, 1982;Marceau et al, 1982;Baribault et al, 1985). Glucocorticoids such as dexamethasone can modulate the proliferation of various cell types and promote the differentiation of various tissues (Guerriero and Florini, 1980;Belanger et al, 1981).…”

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confidence: 99%

Dexamethasone and dimethylsulfoxide as distinct regulators of growth and differentiation of cultured suckling rat hepatocytes

Baribault

Marceau

1986

Journal Cellular Physiology

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Dexamethasone can promote the differentiation of different tissues in vivo while dimethylsulfoxide is a commonly used inducer of differentiation in various tumor cell types in culture. In the present study, the effects of dexamethasone and dimethylsulfoxide on growth and functional activities of cultured differentiating suckling rat hepatocytes stimulated with various combinations of EGF, insulin, and glucagon were evaluated. Hepatocytes stimulated with EGF and either insulin or glucagon entered S phase and mitosis after a lag period of 24 h. These hormonal factors thus provide simple combinations of hepatocyte-growth regulators. Dexamethasone in the presence of EGF and glucagon inhibited the initiation of DNA synthesis and mitosis, but it had no effect on EGF-insulin stimulated cultures. Such a differential effect of dexamethasone was observed at concentrations ranging from 4 nM to 200 microM. alpha-Fetoprotein, albumin, and tyrosine aminotransferase were used as typical markers of hepatocyte differentiation status. Irrespective of the combinations of growth-promoting factors used, dexamethasone inhibited alpha 1-fetoprotein production and maintained albumin production and tyrosine aminotransferase inducibility. In contrast, dimethylsulfoxide at 2% inhibited hepatocyte growth and supported the maintenance of the production of both alpha 1-fetoprotein and albumin, independent of the hormonal growth regulators used. On this basis, dexamethasone and dimethylsulfoxide act as distinct modulators of growth and maturation of cultured differentiating suckling rat hepatocytes.

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“…Likewise Parzefall et al (19) and Tomita et al (20) induced DNA synthesis and mitosis in the presence of sera and McGowan et al (21) and McGowan and Bucher (22) reported DNA synthesis and mitoses in a serum-free medium after plating in serum. In a totally serum-free culture system, Baribault et al (23) found significant mitoses but only in hepatocytes from suckling rats and very few from adult rats. Hasegawa et al (24) reported time lapse cinematography of mitoses in cells from adult rats cultured in a serumfree medium, but only if the calcium concentration was reduced below 0.4 mM and a trypsin inhibitor was used.…”

mentioning

confidence: 99%

Electron microscopic and time lapse studies of mitosis in cultured rat hepatocytes

Sattler¹,

Sawada²,

Sattler³

et al. 1988

Hepatology

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Primary cultures of adult rat hepatocytes in a serum-free medium were observed by time lapse cinematography to proceed through mitotis and cytokinesis. An ultrastructural study of these cultures is presented with electron micrographs of each stage of mitosis and cytokinesis. The cultured hepatocytes begin to enter prophase about 48 hr after plating and proceed through mitosis in approximately 70 min not including cytokinesis. During this time, they remain somewhat flattened and joined to neighboring cells rather than rounding up. Both mononucleate and binucleate hepatocytes proceed through mitosis. Some mononucleate cells do not undergo cytokinesis, resulting in the formation of binucleate cells. In binucleate hepatocytes, both nuclei proceed through prophase simultaneously. Usually a single mitotic spindle with a large metaphase plate containing chromosomes from both nuclei is observed. Cytokinesis frequently occurs in binucleate hepatocytes which have a single mitotic spindle. Some binucleate cells form tripolar or 4-polar metaphase plates. In tripolar metaphases, some cells do not divide, resulting in multinucleate cells, whereas others undergo cytokinesis yielding three mononucleate cells or one binucleate and one mononucleate cell. Two mitotic spindles located perpendicularly to each other with microtubules intertwining have been observed in 4-polar metaphases. In this latter case, no cytokinesis has been observed. This study shows that both mononucleate and binucleate adult rat hepatocytes cultured in a serum-free medium in the presence of epidermal growth factor not only synthesize DNA, but progress through mitosis and often cytokinesis.

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Differential responsiveness of cultured suckling and adult rat hepatocytes to growth‐promoting factors: Entry into s phase and mitosis

Cited by 31 publications

References 34 publications

Differential activity of a tissue-specific extinguisher locus in hepatic and nonhepatic cells.

Differential activity of a tissue-specific extinguisher locus in hepatic and nonhepatic cells.

Dexamethasone and dimethylsulfoxide as distinct regulators of growth and differentiation of cultured suckling rat hepatocytes

Electron microscopic and time lapse studies of mitosis in cultured rat hepatocytes

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