Skip to main content
SpringerLink
Log in

Expression of ACTH-induced corticosteroid biosynthesis in newborn rat adrenocortical cells cultured in serum-free and carrier protein-free medium containing a cholesterol-α-cyclodextrin complex

  • Published:
Cell Biology and Toxicology Aims and scope Submit manuscript

Abstract

Newborn rat adrenocortical cells were successfully cultured in a serum free carrier protein free medium (SPFM) by using α-cyclodextrin as a cholesterol carrier and have expressed corticosteroid biosynthesis in this medium. A stable inclusion complex of cholesterol-α-cyclodextrin with a molar ratio of almost 1 was obtained for a 5 × 10−5 mol/1 α-cyclodextrin concentration. Cell cultures incubated with [4-14C] cholesterol-α-cyclodextrin in SPFM produced, under ACTH stimulation, various 14C labeled steroids with a predominance of corticosterone and 18-hydroxy-11-deoxycorticosterone. As measured by gas chromatography and mass spectrometry, the ratio between corticosteroids (21-hydroxylated steroids) and 20α-reduced steroids produced in SPFM with cholesterol-α-cyclodextrin was equal to 1.8. This corresponds to a value of 3.6 times higher than that found in the serum free medium with cholesterol-albumin. Consequently, the chemically defined SPFM with cholesterol-α-cyclodextrin used in this study is more suitable for corticosteroidogenesis by adrenal cells in culture than a serum free medium with cholesterol-albumin.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

α-CD:

α-cyclodextrin

ACTH:

adrenocorticotropic hormone

20α-dihydroprogesterone:

20α-hydroxy-4-pregnene-3-one

11β-hydroxy-20α-dihydroprogesterone:

11β, 20α-dihydroxy-4-pregnene-3-one

11β-hydroxyprogesterone:

11β-hydroxy-4-pregnene-3,20-dione

C-α-CD:

cholesterol-α-cyclodextrin complex

corticosterone:

11β,21-dihydroxy-4-pregnene-3,20-dione

deoxycorticosterone:

21-hydroxy-4-pregnene-3,20-dione

18-hydroxy-11-deoxycorticosterone:

18,21-dihydroxy-4-pregnene-3,20-dione

18-hydroxy-20α-dihydroprogesterone:

18-20α-dihydroxy-4-pregnene-3-one

18-hydroxyprogesterone:

18-hydroxy4-pregnene-3,20-dione

progesterone:

4-pregnene-3,20-dione

SFM-S:

serum-free medium

SPFM:

serum-free protein-free medium

SSM:

serum supplemented medium

References

  • BARNES, D. and SATO, G. (1980). Serum-free cell cultures: a unifying approach. Cell 22:649–655.

    Google Scholar 

  • BERTOLINO, A., BENSON, A.M. and TALALAY, P. (1979). Activation of γ5-3-ketosteroid isomerase of bovine adrenal microsomes by serum albumins. Biochem. Biophys. Res. Commun. 88:1158–1166.

    Google Scholar 

  • CHESSEBEUF, M. and PADIEU, P. (1984). Rat liver epithelial cell cultures in a serum-free medium: primary cultures and derived cell lines expressing differentiated functions. In Vitro 20:780–795.

    Google Scholar 

  • CHESSEBEUF, M., FISCHBACH, M. and PADIEU, P. (1984). Time course study of L-Tyrosine aminotransferase induction in rat liver cell lines. Cell Biol. Toxicol. 1:31–40.

    Google Scholar 

  • FRANK, D.W., GRAY, J.E. and WEAVER, R.N. (1976). Cyclodextrin nephrosis in the rat. Ann. J. Pathol. 83:367–374.

    Google Scholar 

  • GAMBERT, P., LALLEMANT, C., ARCHAMBAULT, A., MAUME, B.F. and PADIEU, P. (1979). Assessment of serum cholesterol by two methods: gas-liquid chromatography on capillary column and chemical ionization mass fragmentography with isotopic dilution of |3,4−13C| cholesterol as internal standard. J. Chromatogr. Biomed. App. 162:1–6.

    Google Scholar 

  • GWYNNE, J.T. and HESS, B. (1980). The role of high density lipoproteins in rat adrenal cholesterol metabolism and steroidogenesis. J. Biol. Chem. 255:10875–10883.

    Google Scholar 

  • HAMILTON, M.A., McCUNE, R.W. and ROBERTS, S. (1972). Activation of solubilized steroid transforming enzymes of adrenal microsomal origin by serum protein. Endocrinology 54:297–315.

    Google Scholar 

  • HAMMAMI, M., LEGENDRE C. and MAUME, B.F. Induction of corticosteroid pathways by ACTH and high density lipoproteins in newborn rat adrenocortical cells cultured in serum-free medium. Biochim. Biophys. Acta, submitted.

  • KOVANEN, P.T., GOLDSTEIN, J.L., CHAPPELL, D.A. and BROWNN, M.S. (1980). Regulation of low density lipoprotein receptors by adrenocorticotropin in the adrenal gland of mice and rats in vivo. J. Biol. Chem. 255:5591–5598.

    Google Scholar 

  • KRENNING, E.P., DOCTER, R., BERNARD, H.F., VISSER, T.J. and HENNEMANN, G. (1979). The essential role of albumin in the active transport of thyroid hormones into primary cultured rat hepatocytes. FEBS Letters 107:227–230.

    Google Scholar 

  • MANEVIC, E.M., MUZYA, G.I., PROKAZOVA, N.V., MOLOTKOVSKY, J.G., BEZYGLOV, V.V. and BERGELSON, L.D. (1984). Interaction of prostaglandin E1 with human high density lipoproteins. FEBS Letters 173:291–294.

    Google Scholar 

  • McNEIL, P.L., MURPHY, R.F., LANNI, F. and TAYLOR, D.L. (1984). A method for incorporating macromolecules into adherent cells. J Cell Biol 98:1556–1564.

    Google Scholar 

  • MAUME, B.F. and PROST, M. (1973). Maintien des functions de steroidogenese differencice chez des cellules surrenaliennes du rat nouveau-ne maintenues en culture monocouche. C. R. Soc. Biol. 167:1427–1430.

    Google Scholar 

  • MAUME, B.F., PROST, M. and PADIEU, P. (1976). Steroid hormone biosynthesis in adrenal cell cultures from newborn rats. In: Advances in Mass Spectrometry in Biochemistry and Medicine (A. Frigerio and N. Castagnoli, eds), Vol. 1, pp. 525–540, Spectrum Publications, New York.

    Google Scholar 

  • MAUME, B.F., MILLOT, C., MESNIER, D., PATOURAUX, D., DOUMAS, J. and TOMORI, E. (1979). Quantitative analysis of corticosteroids in adrenal cell cultures by capillary column gas chromatography combined with mass spectrometry. J. Chromatogr. 186:581–594.

    Google Scholar 

  • PADIEU, P., MAUME, G., HUSSEIN, N., CHESSEBEUF, M. and TSACONAS, C. (1985). Biosynthesis of sterols and bile acids in rat liver epithelial cell lines. Biochem. Biophys. Acta 833:245–261.

    Google Scholar 

  • PROST, M. and MAUME, B.F. (1974). Hormones steroides de la surrenale de rat: analyse des 18-hydroxycorticosteroides par chromatographie gaz-liquide couplee a la spectrometrie de masse et par fragmentographie de masse. J. Steroid Biochem. 5:133–144.

    Google Scholar 

  • RAMIREZ, L.C., MILLOT, C. and MAUME, B.F. (1982). Sample purification using a C18-reversed-phase cartridge for the quantitative analysis of corticosteroids in adrenal cell cultures by high performance liquid chromatography or gas chromatography-mass spectrometry. J. Chromatogr. 229:267–281.

    Google Scholar 

  • RENSHAW, P.F., JANOFF, A.S. and MILLER, K.W. (1983). On the nature of dilute aqueous cholesterol suspensions. J. Lipid Res. 24:47–51.

    Google Scholar 

  • SAENGER, W. (1980). Cyclodextrin inclusion compounds in research and industry. Angew. Chem. Int. Ed. Engl. 19:344–362.

    Google Scholar 

  • SATOMI, D. and KISHIMOTO, Y. (1981). Change of galactolipids and metabolism of fatty acids in the organotypic culture of myelinating mouse brain. Biochim. Biophys. Acta 666:446–454.

    Google Scholar 

  • SINGH, I. and KISHIMOTO, Y. (1983). Effect of cyclodextrins on the solubilization of lignoceric acid, ceremide, and cerebroside, and on the enzymatic reactions involving these compounds. J. Lipid Res. 16:662–665.

    Google Scholar 

  • YAMANE, I. (1981). Alpha-cyclodextrin, a novel substitute for bovine albumin in serum-free culture of mammalian cells. Proc. Jpn. Acad. 57:385–389.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Biochimie des Interactions Cellulaires, Faculté de Sciences, Dijon, France

    Mohammed Hammami, Gabrielle Maume & Bernard F. Maume

Authors
  1. Mohammed Hammami
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gabrielle Maume
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bernard F. Maume
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hammami, M., Maume, G. & Maume, B.F. Expression of ACTH-induced corticosteroid biosynthesis in newborn rat adrenocortical cells cultured in serum-free and carrier protein-free medium containing a cholesterol-α-cyclodextrin complex. Cell Biol Toxicol 2, 41–52 (1986). https://doi.org/10.1007/BF00117706

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00117706

Key words

Search

Navigation