Secretion of albumin and alpha-foetoprotein by dimethylsulphoxide-stimulated hepatocellular carcinoma cells

Higgins, Paul J.; Darzynkiewicz, Zbigniew; Melamed, Myron R.

doi:10.1038/bjc.1983.221

Cited by 21 publications

(12 citation statements)

References 24 publications

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“…Recent investigations have shown that Me2SO treatment of mouse and rat hepatoma cells results in loss of the ability to clone in soft agar medium (17), increased albumin production with a corresponding decline in y-glutamyl transpeptidase expression (18), increased transferrin production (19), and a decline in the percentage of cells in the diploid G1 population with a corresponding increase in the proportion of cells in either a G2 or tetraploid G1 population (20). To our knowledge, the use of Me2SO to maintain differentiation in normal hepatocytes has not been reported prior to this study.…”

Section: Discussionmentioning

confidence: 99%

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Maintenance of differentiated rat hepatocytes in primary culture.

Isom¹,

Secott²,

Georgoff³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

320

188

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Normal adult rat hepatocytes remained viable and functional for at least 43 days when plated on collagen-coated dishes and fed chemically defined medium supplemented with dimethyl sulfoxide (Me2SO). Hepatocytes isolated by collagenase perfusion and cultured in the presence or absence of Me2SO were (i) examined by light and electron microscopy for morphological changes; (it) analyzed for the production of albumin and other plasma proteins; and (iiM) tested by autoradiography for DNA synthesis. Me2SO-treated cells continued to produce specific plasma proteins during the entire culture period; albumin production was consistently high (11)(12)(13)(14)(15)(16)(17)(18)(19) ,.g/ml of culture medium per 24 hr) from day 2 to at least day 43 after plating. Ultrastructural analyses demonstrated that Me2SO-treated hepatocytes resembled those from intact liver in organization of cytoplasmic organelles and cellular junctions. The optimal concentration for observing the morphological and biochemical effects of Me2SO was 2% (vol/vol). We conclude that supplementation of chemically dermed medium with Me2SO enables maintenance ofdifferentiated hepatocytes in culture for extended periods of time.Dimethyl sulfoxide (Me2SO) is a dipolar aprotic solvent that is active in biological systems (1). Addition of 1-2% (vol/vol) Me2SO to the culture medium of Friend virus-induced murine erythroleukemia (MEL) cells for 4-5 days causes 90% of the cells to express characteristics associated with normal erythroid differentiation, including alterations in morphology (2), induction of a-and P-globin synthesis (3, 4), and loss of the capacity for cell division (5). Me2SO-induced differentiation has also been observed in a human leukemia cell line (6) and in cultured fibrosarcoma (7), neuroblastoma (8), human colon carcinoma (9), human lung cancer (10), and murine embryonal carcinoma (11, 12) cell lines.Past efforts to achieve long-term culture of differentiated normal adult hepatocytes have not been successful. Limited proliferation and maintenance of adult hepatocytes can be achieved by supplementing culture medium with serum from partially hepatectomized animals (13) or by plating hepatocytes on liver extracellular matrix and maintaining them in serum-free hormonally defined medium (14). Proliferation also can be achieved by culturing hepatocytes at low cell density in the presence of insulin and epidermal growth factor (EGF), but maintenance of hepatocyte-specific characteristics requires high density or supplementation with hepatic plasma membrane material (15,16).In the present study, we employed a collagen-coated surface and supplemented the culture medium with Me2SO in an attempt to extend the time in vitro that hepatocytes remain biochemically and morphologically differentiated. The addition of Me2SO had a dramatic effect; hepatocytes retaining morphological and biochemical characteristics of normal liver could be maintained in culture for as long as 43 days. Note that Me2SO, used previously to induce differentiation in tumor cells ...

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Section: Discussionmentioning

confidence: 99%

“…The addition of Me2SO had a dramatic effect; hepatocytes retaining morphological and biochemical characteristics of normal liver could be maintained in culture for as long as 43 days. Note that Me2SO, used previously to induce differentiation in tumor cells (2,(6)(7)(8)(9)(10)(11)(12)(17)(18)(19)(20), is used here to maintain differentiation of a normal cell. …”

mentioning

confidence: 99%

Maintenance of differentiated rat hepatocytes in primary culture.

Isom¹,

Secott²,

Georgoff³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

320

188

View full text Add to dashboard Cite

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“…Treatment of hepatocytes with DMSO has been shown to enhance the expression of hepatocyte differentiationspecific markers [42][43][44] and the expression of HBV [24] in vitro. Figure 6 shows that HBV expression was slightly enhanced by the inclusion of 1% DMSO in standard medium containing 10% FCS.…”

Section: Treatment With Dmsomentioning

confidence: 99%

Optimised Conditions for the Production of Hepatitis B Virus from Cell Culture

Glebe¹,

Berting²,

Broehl³

et al. 2001

Intervirology

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In chronically infected patients, hepatitis B virus (HBV) particles reach numbers as large as >10⁹ genome equivalents (GE)/ml of serum. However, expression of infectious HBV particles in cell culture only yields 10⁵–10⁶ GE/ml, which is insufficient for many studies. HBV transcription and possibly replication is dependent on hepatocyte-specific differentiation. Thus, we tested several cell culture parameters that have been reported to enhance the expression of hepatocyte-specific markers, such as growth on different extracellular matrices, different cell culture media, low concentrations of fetal calf serum (FCS) and the addition of dimethyl sulfoxide (DMSO) to the medium. Lower concentrations of FCS, growth on collagen and inclusion of DMSO in the medium only moderately enhanced HBV production in vitro when applied individually. However, combinations of these parameters optimised cell culture conditions and reproducibly increased the release of HBV particles about 100-fold to titres >10⁸ GE/ml of culture medium.

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“…Exposure of mouse hepatoma cells to dimethyl sulfoxide (DMSO) stimulates albumin and AFP accumulation in the medium (Higgins et al, 1983). Similarly, changes in hepatocyte gene expression occur during treatment of rat and mouse hepatoma Correspondence: T. Nakagawa.…”

mentioning

confidence: 99%

Effects of sodium n-butyrate on alpha-fetoprotein and albumin secretion in the human hepatoma cell line PLC/PRF/5

Nakagawa¹,

Nakao²,

Matsui³

et al. 1985

Br J Cancer

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Summary The in vitro effects of sodium n-butyrate (butyrate) on the growth, morphology and secretion of alpha-fetoprotein (AFP) and albumin by the human heptoma cell line PLC/PRF/5 were studied. Butyrate caused a marked reduction in the growth rate, colony forming efficiency in soft agar and de novo synthesis of DNA as well as remarkable morphological changes including cell enlargement, flattening and a decreased number of nucleoli. Secretion of AFP was reduced during culture with butyrate, while that of albumin was increased. The requirement of de novo protein synthesis for the increase in albumin and decrease of AFP by butyrate was demonstrated by inhibition studies with cycloheximide. These results suggest that butyrate caused the hepatoma cells to acquire in vitro properties that are considered to be more consistent with normal liver cells.

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Secretion of albumin and alpha-foetoprotein by dimethylsulphoxide-stimulated hepatocellular carcinoma cells

Cited by 21 publications

References 24 publications

Maintenance of differentiated rat hepatocytes in primary culture.

Maintenance of differentiated rat hepatocytes in primary culture.

Optimised Conditions for the Production of Hepatitis B Virus from Cell Culture

Effects of sodium n-butyrate on alpha-fetoprotein and albumin secretion in the human hepatoma cell line PLC/PRF/5

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