Skip to main content
SpringerLink
Log in

Gonadotropes in a novel rat pituitary tumor cell line, RC-4B/C. Establishment and partial characterization of the cell line

  • Regular Papers
  • Published:
In Vitro Cellular & Developmental Biology Aims and scope Submit manuscript

Summary

An epithelial cell line (RC-4B/C) was established from a pituitary adenoma obtained from a 3-yr-old (ACI/fMai × F344/fMai)F1 male rat. Before Year 5 in vitro, RC-4B/C cells could not be viably recovered from cryogenic storage. Recovery of viable cells from cryogenic storage in Year 5 was associated with a more transformed phenotype, including the appearance of endogenous C-type rat retroviral particles. The ultrastructural appearance of the cells was similar to that of differentiated anterior pituitary cell; the cultured cells contained numerous, electron dense, secretory granules, Golgi complexes, and extended arrays of rough endoplasmic reticulum. Immunocytochemical study showed that all cell types present in the rat anterior pituitary gland were present in the cell line. The percentage of luteinizing hormone beta (LHβ) cells in the cell line was higher (19.9%) and that of growth hormone cells was lower (12.2%) than in normal male rat pituitary, whereas the cell line contained a comparable percentage of follicle stimulating hormone beta (FSHβ), prolactin (PRL), ACTH, and thyrotropin beta cells. Radioimmunoassay data demonstrated the PRL content of the cells was comparable to that of normal male rat pituitary gland, whereas the content of LH and FSH was 70- and 800-fold lower, respectively. Assay of specific receptor sites for gonadotropin releasing hormone (GnRH) using Scatchard plots of the data established the RC-4B/C cells contained GnRH receptor sites of the same affinity as in the pituitary gland, but of twofold lower capacity. These data suggest the RC-4B/C cell line warrants further study as a model for the induction and maintenance of the gonadotropic function of the pituitary gland.

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

References

  1. Ahmed, M.; Korol, W.; Larson, D., et al. Transformation of rat mammary cell cultures by R-35 virus isolated from spontaneous rat mammary adenocarcinoma. JNCI 48:1077–1083; 1972.

    PubMed  CAS  Google Scholar 

  2. Bedigian, H. G.; Taylor, B. A.; Meier, H. Expression of murine leukemia virus in the highly lymphomatous BXH-2 recombinant inbred strain. J. Virol. 39:632–640; 1981.

    PubMed  CAS  Google Scholar 

  3. Berkvens, J. M.; Nesselrooy, J. H. J.; Kroes, R. Spontaneous tumors in the pituitary gland of old wistar rats. A morphologic and immunocytochemical study. J. Pathol. 130:179–191; 1980.

    Article  Google Scholar 

  4. Buonassissi, V.; Sato, G.; Cohen, A. L. Hormone-producing cultures of adrenal and pituitary tumor origin. Proc. Natl. Acad. Sci. USA 48:1184–1190; 1962.

    Article  Google Scholar 

  5. Chattopadhyay, S. K.; Lander, M. R.; Rands, E., et al. The structure of endogenous murine leukemia virus DNA in mouse genomes. Proc. Natl. Acad. Sci. USA 77:5774–5778; 1980.

    Article  PubMed  CAS  Google Scholar 

  6. Chen, T. R. In situ detection of mycoplasm contamination in cell cultures by fluorescent Hoechst 33258 stain. Exp. Cell Res. 104:255–262; 1977.

    Article  PubMed  CAS  Google Scholar 

  7. Childs, G. V. The use of multiple methods to validate immunocytochemical stains. J. Histochem. Cytochem. 31:168–176; 1983.

    PubMed  CAS  Google Scholar 

  8. Dada, M. O.; Campbell, G. T.; Blake, C. A. Pars distalis cell quantification in normal adult male and female rats. J. Endocrinol. 101:87–94; 1984.

    PubMed  CAS  Google Scholar 

  9. Davisson, M. T.; Akeson, E. C. An improved method for preparing G-banded chromosomes from mouse peripheral blood. Cytogenet. Cell Genet. 45:70–74; 1987.

    PubMed  CAS  Google Scholar 

  10. Denef, C.; Hautekeete, E.; Rubin, L. A specific population of gonadotrophs purified from immature female rat pituitary. Science 194:848–851; 1976.

    Article  PubMed  CAS  Google Scholar 

  11. Dubois, M. P. Mise en évidence par immunofluorescence des cellules somatotropes et des cellules à prolactine dans l'hypophyse foetale des bovins. C. R. Acad. Sci. Paris, Série D, 272:433–435; 1971.

    CAS  Google Scholar 

  12. Dubois, M. P. Les cellules corticotropes de l'hypophyse des bovins et porcins. Mise en évidence par immunofluorescence et caractères cytologiques. Ann. Biol. Anim. Biochem. Biophys. 11:584–624; 1971.

    Google Scholar 

  13. Freshney, R. I. Culture of animal cells. A manual of basic technique. 2nd ed. New York: Alan R. Liss, Inc.;1987.

    Google Scholar 

  14. Graham, R. C.; Karnovsky, M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney. Ultrastructural cytochemistry by a new technique. J. Histochem. Cytochem. 14:291–302; 1966.

    PubMed  CAS  Google Scholar 

  15. Hopkins, C. R.; Farquhar, M. G. Hormone secretion by cells dissociated from rat anterior pituitaries. J. Cell. Biol. 59:276–303; 1973.

    Article  CAS  PubMed  Google Scholar 

  16. Hyde, C. L.; Childs, G.; Wahl, L. M. et al. Preparation of gonadotroph-enriched cell population from adult rat anterior pituitary cells by centrifugal elutriation. Endocrinology 111:1421–1423; 1982.

    PubMed  CAS  Google Scholar 

  17. Ibrahim, S. N.; Moussa, S. M.; Childs, G. V. Morphometric studies of rat anterior pituitary cells after gonadectomy: correlation of changes in gonadotropes with the serum levels of gonadotropins. Endocrinology 119:629–637; 1986.

    PubMed  CAS  Google Scholar 

  18. Jameson, J. L.; Lindell, C. M.; Habener, J. F., et al. Abnormal transcription of luteinizing hormone beta-subunit messenger ribonucleic acid in an alpha-subunit secreting pituitary adenoma. Clin. Res. 33:600A; 1985.

    Google Scholar 

  19. Joyner, A.; Keller, G.; Phillips, R. A., et al. Retro-virus transfer of a bacterial gene into mouse hematopoietic progenitor cells. Nature 305:556–558; 1983.

    Article  PubMed  CAS  Google Scholar 

  20. Khar, A.; Debeljuk, L.; Jutisz, M. Biosynthesis of gonadotropins by rat pituitary cells in culture and in pituitary homogenates. Effect of gonadotropin releasing hormone. Mol. Cell. Endocrinol. 12:53–65; 1978.

    Article  PubMed  CAS  Google Scholar 

  21. Kovacs, K.; Horvath, E. Pathology of pituitary adenomas. In: Givens, J. R., ed. Hormone secreting pituitary tumors. Chicago: Year Book Publ. Inc.; 1982:97–119.

    Google Scholar 

  22. Labarca, C.; Paigen, K. A simple, rapid and sensitive DNA assay procedure. Anal. Biochem. 102:344–352; 1980.

    Article  PubMed  CAS  Google Scholar 

  23. Leiter, E. H.; Malinoski, F. J.; Eppig, J. J. An epithelial cell line with chronic polyoma infection established from a spontaneous mouse pancreatic adenocarcinoma. Cancer Res. 38:969–977; 1978.

    PubMed  CAS  Google Scholar 

  24. Leong, D. A.; Lau, S. K.; Sinha, Y. N., et al. Enumeration of lactotropes and somatotropes among male and female pituitary cells in culture: evidence in favor of a mammosomatotrope subpopulation in the rat. Endocrinology 116:1371–1378; 1985.

    PubMed  CAS  Google Scholar 

  25. Liposits, Z.; Gores, T.; Domany, S., et al. Silver intensification of the diaminobenzidine end product in histochemistry and immunohistology. In: Endroczi, E.; Angelucci, I.; Scapagnini, W., et al. eds. Neuropeptides, neurotransmitters and regulation of endocrine processes. Budapest: Academiai Kiado; 1983:383–398.

    Google Scholar 

  26. Maurer, R. A. Use of recombinant DNA procedure to study mechanisms controlling synthesis of hypophyseal hormones. Biol. Reprod. 28:27–35; 1983.

    Article  PubMed  CAS  Google Scholar 

  27. McComb, D. J.; Kovacs, K.; Beri, J., et al. Pituitary adenomas in old Sprague-Dawley rats: a histologic, ultrastructural and immunocytochemical study. JNCI 73:1143–1166; 1984.

    PubMed  CAS  Google Scholar 

  28. McComb, D. J.; Kovacs, K.; Beri, J., et al. Pituitary gonadotroph adenomas in old Sprague-Dawley rats. J. Submicrosc. Cytol. 17:517–530; 1985.

    PubMed  CAS  Google Scholar 

  29. Nakane, P. K.; Pierce, G. B. Enzyme labelled antibodies preparation and application for antigen localization. J. Histochem. Cytochem. 14:929–931; 1966.

    PubMed  CAS  Google Scholar 

  30. Noël, N.; Bérault, A.; Théoleyre, M., et al. Pituitary GnRH receptor content and LH release from rat pituitary cells in culture: effect of castration in the male rat. Neuroendocrinol. Lett. 9:231–236; 1987.

    Google Scholar 

  31. Shiino, M.; Maruyama, T.; Tanaka, T. Gonadotroph-rich cell lines derived from pituitary clonal cells (2A8) grafted under the kidney capsule. Cell Tissue Res. 201:93–99; 1979.

    Article  PubMed  CAS  Google Scholar 

  32. Snyder, P. J. Gonadotroph cell adenomas of the pituitary. Endocrine Rev. 6:552–563; 1985.

    CAS  Google Scholar 

  33. Snyder, P. J.; Johnson, J.; Muzyka, R. Abnormal secretion of glycoprotein α-subunit and follicle-stimulating hormone (FSH) β-subunit in men with pituitary adenomas and FSH hypersecretion. J. Clin. Endocr. Metab. 51:579–584; 1980.

    PubMed  CAS  Google Scholar 

  34. Starzec, A.; Jutisz, M.; Counis, R. Cyclic adenosine monophosphate and phorbol ester, like gonadotropin-releasing hormone, stimulate the biosynthesis of luteinizing hormone polypeptide chains in a nonadditive manner. Mol. Endocrinol. 3:618–624; 1989.

    Article  PubMed  CAS  Google Scholar 

  35. Tashjian, A. H., Jr.; Yasumura, Y.; Levine, L., et al. Establishment of clonal strains of rat pituitary tumor cells that secrete growth hormone. Endocrinology 82:342–352; 1968.

    PubMed  CAS  Google Scholar 

  36. Tashjian, A. H., Jr.; Bancroft, F. C.; Levine, L. Production of both prolactin and growth hormone by clonal strains of rat pituitary tumor cells. J. Cell. Biol. 47:61–70; 1970.

    Article  PubMed  CAS  Google Scholar 

  37. Teich, N. Taxonomy of retroviruses. In: Weiss, R.; Teich, N.; Varmus, H., et al. eds. RNA Tumor Viruses. New York: Cold Spring Harbor Laboratory; 1982:25–208.

    Google Scholar 

  38. Tixier-Vidal, A.; Kerdelhué, B.; Jutisz, M. Kinetics of release of luteinizing hormone (LH) and follicle stimulating hormone (FSH) by primary cultures of dispersed rat anterior pituitary cells. Chronic effect of synthetic LH and FSH releasing hormone. Life Sci. 12:499–509; 1973.

    Article  CAS  Google Scholar 

  39. Ulrich, K.; Briand, P. Treatment ofMycoplasma hyorrhinis contaminated tissue cultures with a mixture of antibiotics. Acta Pathol. Microbiol. Scand., Sect. B. 85:347–349; 1977.

    Google Scholar 

  40. Vale, W.; Grant, G.; Amoss, M., et al. Culture of enzymatically dispersed anterior pituitary cells: functional validation of a method. Endocrinology 91:562–572; 1972.

    Article  PubMed  CAS  Google Scholar 

  41. Wide, L.; Lundberg, P. O. Hypersecretion of an abnormal form of follicle-stimulating hormone associated with suppressed luteinizing hormone secretion in a woman with a pituitary adenoma. J. Clin. Endocrinol. & Metab. 53:923–930; 1981.

    Article  CAS  Google Scholar 

  42. Zamboni, L.; De Martino, C. Buffered picric acid-formaldehyde: a new rapid fixative for electron microscopy. J. Cell. Biol. 35:148a; 1967.

    Google Scholar 

Download references

Author information

Author notes

  1. Jolanta Polkowska

    Present address: Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, 05110 Jablonna near, Warsaw, Poland

Authors and Affiliations

  1. Laboratoires des Hormones Polypeptidiques, CNRS, 91198, Gif sur yvette, France

    Ilse Hurbain-Kosmath, Annette Berault, Nadine Noel, Jolanta Polkowska, Anne Bohin & Marian Jutisz

  2. The Jackson Laboratory, 04609, Bar Harbor, Maine

    Edward H. Leiter, Wesley G. Beamer, Hendrick G. Bedigian, Muriel T. Davisson & David E. Harrison

Authors
  1. Ilse Hurbain-Kosmath
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Annette Berault
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nadine Noel
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jolanta Polkowska
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Anne Bohin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Marian Jutisz
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Edward H. Leiter
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Wesley G. Beamer
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Hendrick G. Bedigian
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Muriel T. Davisson
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. David E. Harrison
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported in part by grants DK-17631 (E.H.L.), CA-24145 (W.G.B.), CA-31102 (H.G.B.), AG-01753 (D.E.H.) and HD-1778 (M.T.D.) from the National Institutes of Health, Bethesda, MD, and by a grant from the Association pour la Recherche sur le Cancer, France (M.J.). The NIH is not responsible for the contents of this publication nor do the contents necessarily represent the official views of that agency.

Jolanta Polkowska was a recipient of a Foundation Simone et Cino del Duca grant.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hurbain-Kosmath, I., Berault, A., Noel, N. et al. Gonadotropes in a novel rat pituitary tumor cell line, RC-4B/C. Establishment and partial characterization of the cell line. In Vitro Cell Dev Biol 26, 431–440 (1990). https://doi.org/10.1007/BF02624084

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02624084

Key words

Search

Navigation