Skip to main content
SpringerLink
Log in

Sertoli cells of adult rats “in vitro”

A morphologic and functional study

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

A cell line obtained from isolated seminiferous tubules of adult rat testis has been studied “in vitro” over a period of 35 days.

Light and electron microscopic studies performed from hour 2 to the end of culture have shown the presence of a monomorphic cell population. After 5–6 days of culture the cells formed a monolayer. The cytoplasm of the cells contained numerous lipid bodies and produced numerous projections. The nucleus showed several indentations and one or more nucleoli. From the 9th to the 15th day of culture the cells developed a large amount of endoplasmic reticulum, Golgi apparatus and aggregates of electron dense granules. From the 20th to 40th day the cell cultures progressively degenerated.

Immunochemical analysis of the culture medium revealed the presence of estradiol-17β, which reached its maximum production rate from the 8th day to the 18th day of culture. Corresponding to cell involution estradiol concentration underwent a rapid decrease.

On the basis of morphological and biochemical data the cells could be considered Sertoli cells.

This work was supported by Grants n.∘ 74.00155.04 and n.∘ 75.01224.04 from the Consiglio Nazionale delle Ricerche (C.N.R.), Rome, Italy, and by Istituto di Ricerca F. Angelini, Rome, Italy

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

  • Beall, D.: The isolation of α-oestradiol and oestrone from horse testes. Bioehem. J. 34, 1293–1298 (1940)

    Google Scholar 

  • Bell, J.B.G., Vinson, G.P., Lacy, D.: Studies on the structure and function of the mammalian testis. III. In vitro steroidogenesis by the seminiferous tubules of rat testis. Proc. roy. Soc. B 176, 433–443 (1971)

    Google Scholar 

  • Berthrong, M., Goodwin, W.E., Scott, W.W.: Estrogen production by the testis. J. clin. Endocr. 9, 579–592 (1949)

    Google Scholar 

  • Brinkmann, A.O., Mulder, E., Lamers-Stahlhofen, G.J.M., Mechielsen, M.J., Molen, H.J. van der: On oestradiol receptor in rat testis interstitial tissue. FEBS Lett. 26, 301–305 (1972)

    Google Scholar 

  • Brökelmann, J.: Fine structure of germ cells and Sertoli cells during the cycle of the seminiferous epithelium in the rat. Z. Zellforsch. 59, 820–850 (1963)

    Google Scholar 

  • Brouha, L., Simonnet, H.: Action d'extraits orchitiques liposolubles sur le tractus génital femelle. C.R. Soc. Biol. (Paris) 99, 41–42 (1928)

    Google Scholar 

  • Budd, J.W.: Gynecomastia associated with interstitial cell tumor of the testis. Amer. J. Path. 13, 660–661 (1937)

    Google Scholar 

  • Cameron, D., Markwald, R., Worthington, W.D.: Ultrastructural and histochemical effects of FSH on Sertoli cells of prepuberal rats. Anat. Rec. 172, 284 (1972)

    Google Scholar 

  • Castro, A.E., Alonso, A., Mancini, R.E.: Localization of follicle stimulating and luteinizing hormones in the rat testis using immunohistological tests. J. Endocr. 52, 129–136 (1972)

    Google Scholar 

  • Castro, A.E., Seiguer, A.C., Mancini, R.E.: Electron microscopic study of the localization of labelled gonadotropins in the Sertoli and Leydig cells of the rat testis. Proc. Soc. exp. Biol. (N.Y.) 133, 582–586 (1970)

    Google Scholar 

  • Christensen, A.K., Mason, N.R.: Comparative ability of seminiferous tubules and interstitial tissue of rat testis to synthesize androgens from progesterone-4-C14 “in vitro”. Endocrinology 74, 646–656 (1965)

    Google Scholar 

  • Cooke, B.A., Jong, F.H. de, Molen, H.J. van der, Rommerts, F.F.G.: Endogenous testosterone concentrations in rat testis interstitial tissue and seminiferous tubule during “in vitro” incubations. Nature (Lond.) New Biol. 237, 255–256 (1972)

    Google Scholar 

  • Courot, M.: Effect of gonadotropins on the seminiferous tubules of the immature testis. In: The human testis (E. Rosenberg and C.A. Paulsen, eds.), pp. 355. New York-London: Plenum Press 1970

    Google Scholar 

  • Courot, M., Hochereau de Reviers, M.T., Ortavant, R.: Spermatogenesis. In: The testis (A.D. Johnson, W.R. Gomes and N.L. Van Demark, eds.), pp. 339. New York-London: Academic Press 1970

    Google Scholar 

  • Cunningham, B., May, J., Gordon, S.: The presence of estrogenic hormone(s) in testicular material. Proc. Soc. exp. Biol. (N.Y.) 49, 130–132 (1942)

    Google Scholar 

  • Davis, J.C.: Isolation of Sertoli cells from 15 day old rat testes by equilibrium centrifugation. J. Cell Biol. 67, 86A (1975)

  • De Martine, C.: Aspetti ultrastrutturali del testicolo umano normale e patologico. In: Gli ipogonadismi maschili (C. Conti and A. Fabbrini, eds.). Roma: Edizioni L. Pozzi 1968

    Google Scholar 

  • De Martino, C., Marcante, M.L., Floridi, A., Citro, G., Cantafora, A., Bellocci, M., Natali, P.G.: Biochemical and morphological studies “in vitro” of presumptive sertoli cells of adult rats. X. Congr. It. Microsc. Elettr. Ostuni 1–4 October 1975. J. submicrosc. Cytol. 8, 250A (1976)

  • De Martino, C., Stefanini, M., Floridi, A., Marcante, M.L., Agrestini, A., Morelli, M.: Morphological and functional modification of mitochondria in the rat germ cells. J. submicr. Cytol. 6, 115–116 (1974)

    Google Scholar 

  • Dodds, E.C., Greenwood, A.W., Gallimore, E.J.: Note on a water-soluble active principle isolated from the mammalian testis and urine, and its relation to oestrin. Lancet 1930I, 683–685

  • Dorfman, R.I., Gallagher, T.F., Kogh, F.C.: The nature of the estrogenic substance in human male urine and bull testis. Endocrinology 19, 33–41 (1935)

    Google Scholar 

  • Dorrington, J.H., Armstrong, D.T.: Follicle-stimulating hormone stimulates estradiol-17β synthesis in cultured Sertoli cells. Proc. nat. Acad. Sci. (Wash.) 72, 2677–2681 (1975)

    Google Scholar 

  • Dorrington, J.H., Fritz, I.B.: Effects of gonadotropins on cyclic AMP production by isolated seminiferous tubule and interstitial cell preparations. Endocrinology 94, 395–403 (1974)

    Google Scholar 

  • Dorrington, J.H., Fritz, I.B.: Cellular localization of 5 α-reductase and 3 α-hydroxysteroid dehydrogenase in the seminiferous tubule of the rat testis. Endocrinology 96, 879–889 (1975)

    Google Scholar 

  • Dorrington, J.H., Roller, N., Fritz, I.B.: The effects of FSH on cell preparations from the rat testis. In: Hormone binding and target cell activation of the testis (M. Dufau and A.R. Means, eds.), Vol. I, pp. 237. New York-London: Plenum Press 1974

    Google Scholar 

  • Dorrington, J.H., Vernon, R.G., Fritz, I.B.: The effect of gonadotrophins on the 3′, 5′-AMP levels of seminiferous tubules. Biochem. biophys. Res. Commun. 46, 1523–1528 (1972)

    Google Scholar 

  • Ellis, L.C., Kämpen, K.R. van: Androgen synthesis and metabolism by rat testicular minced and teased-tubular preparation after 450 R of whole-body X-irradiation. Radiat. Res. 48, 146–154 (1971)

    Google Scholar 

  • Fellner, O.O.: Ueber die Wirkung des Placentar- und Hodenlipoids auf die männlichen und weiblichen Sexualorgane. Pflügers Arch. ges. Physiol. 189, 199–214 (1921)

    Google Scholar 

  • Floridi, A., Citro, G., Sega, E., Natali, P.G.: Quantitation of estradiol receptors employing a sensitive radioimmunoassay. Immun. Commun. 4, 603–615 (1976)

    Google Scholar 

  • Fritz, I.B., Kopec, B., Lam, K., Vernon, R.G.: Effects of FSH on levels of androgen binding protein in the testis. In: Hormone binding and target cell activation of the testis (M. Dufau and A.R. Means eds.), Vol. I, pp. 311. New York-London: Plenum Press 1974

    Google Scholar 

  • Greulich, W.W., Burford, T.H.: Testicular tumors associated with mammary, prostatic, and other changes in cryptorchid dogs. Amer. J. Cancer 28, 496–511 (1936)

    Google Scholar 

  • Hall, P.F., Irby, D.C., Kretser, D.M. de: Conversion of cholesterol to androgen by rat testis: comparison of interstitial cells and seminiferous tubules. Endocrinology 84, 488–496 (1969)

    Google Scholar 

  • Huggins, C., Eichelberger, L.: Studies on tumors of the testis. I. Water and electrolyte content of testicular tumors and of normal cryptorchid, and estrogenized testis. Cancer Res. 4, 447–452 (1944)

    Google Scholar 

  • Huggins, C., Moulder, P.V.: Estrogen production by Sertoli cell tumors of the testis. Cancer Res. 5, 510–514 (1945)

    Google Scholar 

  • Hunt, V.C., Budd, J.W.: Gynecomastia associated with interstitial cell tumor of the testicle. J. Urol. (Baltimore) 42, 1242–1250 (1939)

    Google Scholar 

  • Innes, J.R.M.: Neoplastic diseases of the testis in animals. J. Path. Bact. 54, 485–498 (1942)

    Google Scholar 

  • Jong, F.H. de, Hey, A.H., Molen, H.J. van der: Effect of gonadotrophins on the secretion of oestradiol17- β and testosterone by the rat testis. J. Endocr. 57, 277–284 (1973)

    Google Scholar 

  • Kodani, M., Kodani, K.: The in vitro cultivation of mammalian Sertoli cells. Proc. nat. Acad. Sci. (Wash.) 56, 1200–1206 (1966)

    Google Scholar 

  • Kuehl, F.A., Patanelli, D.J., Tarnoff, J., Humes, J.L.: Testicular adenylcyclase: stimulation by the pituitary gonadotrophins. Biol. Reprod. 2, 154–163 (1970)

    Google Scholar 

  • Lacy, D.: Androgen dependency of spermatogenesis and the physiological significance of steroid metabolism “in vitro” by the seminiferous tubules. In: The endocrine function of the human testis (V.H.T. James, M. Serio and L. Martini, eds.), Vol. I, pp. 493. New York-London: Academic Press 1973

    Google Scholar 

  • Lacy, D., Bars, J.J., Bell, J.B.G., Callius, P.M., Fyson, P., Pettitt, A.J., Pudney, J.: Steroid metabolism by the seminiferous tubules of mammal and its biological and medical significance. In: Endocrinology-Excerpta Medica Foundation, pp. 838, International Congress Series No. 273, 1973

  • Lacy, D., Pettitt, A.J.: Sites of hormone production in the mammalian testis and their significance in the control of male fertility. Brit. med. Bull. 26, 87–91 (1970)

    Google Scholar 

  • Laqueur, F., Jongh, S.F. de: A female (sexual) hormone. J. Amer. med. Ass. 91, 1169–1172 (1928)

    Google Scholar 

  • Leblond, C.P., Clermont, Y.: Definition of the stage of the cycle of the seminiferous epithelium. Ann. N.Y. Acad. Sci. 55, 548–573 (1952)

    Google Scholar 

  • Lyon, M.F., Glenister, P.H., LynnLamoreux, M.: Normal spermatozoa from androgen-resistent germ cells of chimaeric mice and the role of androgen in spermatogenesis. Nature (Lond.) 258, 620–622 (1975)

    Google Scholar 

  • Mancini, R.E.: Effect of gonadotropin preparations and of urinary FSH and LH on human spermatogenesis. In: The regulation of mammalian reproduction (S.J. Segal, R. Croizier, P.A. Corfman and P.G. Condliffe, eds.), pp. 151. Springfield: Ch.C. Thomas 1973

  • Mancini, R.E., Castro, A.E., Seiguer, A.C.: Histologic localization of follicle stimulating and luteinizing hormones in the rat testis. J. Histochem. Cytochem. 15, 516–525 (1967)

    Google Scholar 

  • Means, A.R.: Early effects of FSH upon testicular metabolism. In: The human testis (E. Rosenberg and A.C. Paulsen, eds.), pp. 301. New York-London: Plenum Press 1970

    Google Scholar 

  • Means, A.R.: Concerning the mechanism of FSH action: rapid stimulation of testicular synthesis of nuclear RNA. Endocrinology 89, 981–989 (1971)

    Google Scholar 

  • Means, A.R.: Early sequence of biochemical events in the action of follicle stimulating hormone on the testis. Life Sci. 15, 371–389 (1974)

    Google Scholar 

  • Means, A.R., MacDougall, E., Soderling, T.R., Corbin, J.D.: Testicular adenosine-3′, 5′-monophosphate-dependent protein kinase. J. biol. Chem. 249, 1231–1238 (1974)

    Google Scholar 

  • Means, A.R., Vaitukaitis, J.: Peptide hormone “receptors”: specific binding of 3H-FSH to testis. Endocrinology 90, 39–46 (1972)

    Google Scholar 

  • Molen, H.J. van der, Brinkmann, A.O., Cooke, B.A., Jong, F.H. de, Mulder, E., Rommerts, F.F.G., Vusse, G.J. van der: Biochemical functions of isolated interstitial tissue and seminiferous tubules from rat testis. In: The endocrine function of the human testis (V.H.T. James, M. Serio and L. Martini, eds.), Vol. I, pp. 535. New York-London: Academic Press 1973

    Google Scholar 

  • Molen, H.J. van der, Bruijn, H.W.A. de, Cooke, B.A., Jong, F.H. de, Rommerts, F.F.G.: Regulation of the production of testicular steroids. In: The endocrine function of the human testis (V.H.T. James, M. Serio and L. Martini, eds.), Vol. I, pp. 459. New York-London: Academic Press 1973

    Google Scholar 

  • Monaschkin, G.B.: Gynäkomastie und Hodentumor. Beitrag zur Frage über die sexualorganischen Wechselbeziehungen. Z. Urol. 20, 8–19 (1926)

    Google Scholar 

  • Mulder, E., Brinkmann, A.O., Lamers-Stahlofen, G.J.M., Molen, H.J. van der: Binding of oestradiol by the nuclear fraction of rat testis interstitial tissue. FEBS Lett. 31, 131–136 (1973)

    Google Scholar 

  • Murad, F., Strauch, B.S., Vaughan, M.: The effects of gonadotrophins on testicular adenylcyclase. Biochim. biophys. Acta (Amst.) 177, 591–598 (1969)

    Google Scholar 

  • Murphy, H.D.: Sertoli cell stimulation following intratesticular injection of FSH in the hypophysectomized rat. Proc. Soc. exp. Biol. (N.Y.) 118, 1202–1205 (1965)

    Google Scholar 

  • Nation, E.F., Edmondson, H.A., Hammack, R.W.: Interstitial cell tumors of the testis. Report of three new cases. Arch. Surg. 48, 415–423 (1944)

    Google Scholar 

  • Overton, J.: The fate of desmosomes in trypsinized tissue. J. exp. Zool. 168, 203–214 (1968)

    Google Scholar 

  • Parvinen, M., Niemi, M.: Distribution and conversion of exogenous cholesterol and sex steroids in the seminiferous tubules and interstitial tissue of rat testis. Steroidologia 2, 129–137 (1971)

    Google Scholar 

  • Reddy, K.J., Svoboda, D.J.: Alteration in rat testes due to an antispermatogenic agent. Light and electron microscopic study. Arch. Path. 84, 376–392 (1967)

    Google Scholar 

  • Regaud, C.: Études sur la structure des tubes séminifères et sur la spermatogénèse chez les mammifères. Arch. d'Anat. Micr. 4, 101–156 (1901)

    Google Scholar 

  • Sertoli, E.: Sulla struttura dei canalicoli seminiferi in rapporto allo sviluppo dei nemaspermi. Arch. Sci. med. 2, 197–246 (1877)

    Google Scholar 

  • Stefanini, M., Ziparo, E., Agrestini, A.: Culture “in vitro” di cellule del Sertoli da testicolo di ratto adulto. Personal comunication 1975

  • Steinberger, E., Steinberger, A., Fisher, M.: Study of spermatogenesis and steroid metabolism in culture of mammalian testes. Recent Progr. Hormone Res. 26, 547–588 (1970)

    Google Scholar 

  • Teilum, G.: Estrogen producing Sertoli cell tumors of human testis and ovary: homologous ovarian and testicular tumors. J. clin. Endocr. 9, 301–318 (1949)

    Google Scholar 

  • Tsang, W.N., Collins, P.M., Lacy, D.: Steroid metabolism by the seminiferous tubules “in vitro” and spermatogenesis in developing rat. J. Reprod. Fertil. 34, 513–517 (1973)

    Google Scholar 

  • Tung, P.S., Dorrington, J.H., Fritz, I.B.: Structural changes induced by follicle-stimulating hormone or dibutyryl cyclic AMP on Sertoli cells in culture. Proc. nat. Acad. Sci. (Wash.) 72, 1838–1842 (1975)

    Google Scholar 

  • Welsh, M.J., Wiebe, J.P.: Rat Sertoli cells: a rapid method for obtaining viable cells. Proc. Can. Fed. Biol. Soc. 17, 44 A (1974)

    Google Scholar 

  • Welsh, M.J., Wiebe, J.P.: Rat Sertoli cells: a rapid method for obtaining viable cells. Endocrinology 96, 618–624 (1975)

    Google Scholar 

  • Wrobel, K.H., Kühnel, W.: Die Histotopik einiger Oxydoreduktasen im Hoden von Hund und Katze. Histochemie 10, 208–215 (1967)

    Google Scholar 

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

  • Zondek, B.: Mass excretion of oestrogenic hormone in the urine of the stallion. Nature (Lond.) 133, 209–210 (1934)

    Google Scholar 

  • Zuckerman, S., Groome, J.R.: The aetiology of benign enlargement of the prostate in the dog. J. Path. Bact. 44, 113–124 (1937)

    Google Scholar 

  • Zuckerman, S., McKeown, T.: The canine prostate in relation to normal and abnormal testicular changes. J. Path. Bact. 46, 7–19 (1938)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Regina Elena Institute for Cancer Research, Rome, Italy

    C. De Martino, M. L. Marcante, A. Floridi, G. Citro, M. Bellocci, A. Cantafora & P. G. Natali

  2. Istituto Superiore di Sanità, Rome, Italy

    A. Cantafora

Authors
  1. C. De Martino
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. L. Marcante
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. Floridi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. G. Citro
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. Bellocci
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A. Cantafora
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. P. G. Natali
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

The excellent technical assistance of Miss Laura Vassallo, Daniela Venturini and Mr. Massimo Rosati and Mario Termine is deeply appreciated

Rights and permissions

Reprints and permissions

About this article

Cite this article

De Martino, C., Marcante, M.L., Floridi, A. et al. Sertoli cells of adult rats “in vitro”. Cell Tissue Res. 176, 69–90 (1977). https://doi.org/10.1007/BF00220345

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00220345

Key words

Search

Navigation