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Establishment of a rat Sertoli cell line that displays the morphological and some of the functional characteristics of the native cell

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Abstract

Primary rat Sertoli cells are widely used as a model for mechanistic and toxicological studies, since they are often the target of toxicants in vivo. However, their isolation from testicular homogenates is tedious and requires the regular use of numerous immature animals. It is therefore of great interest to have available established cell lines that are usable in vitro for unlimited periods and closely similar to native cells. To this end, we have established a line of Wistar rat Sertoli cells (SerW3) by immortalization of fresh primary cells with the T antigens of the Simian virus (SV40). When plated on Matrigel, this cell line presents many of the functional characteristics of Sertoli cells in vivo. In addition, they are sensitive to cisplatin and secrete transferrin, although they do not show a clear response to follicle-stimulating hormone. They also present many morphological similarities, including the presence of tight junctions which mimic the natural epithelial barrier. Like Sertoli cells in vivo, they show extensive phagocytic activity. Finally, they display all the characteristics of immortalized, but not transformed, cells, i.e., topo-inhibition and apoptosis at confluence or under serum deprivation.

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References

  • Aisen P. Iron transport and storage proteins. Annu Rev Biochem. 1980;49:357–93.

    Google Scholar 

  • Bleil JD, Bretscher MS. Transferrin receptor and its recycling in HeLa cells. EMBO J. 1982;3:351–5.

    Google Scholar 

  • Boudreau N, Sympson CJ, Werb Z, Bissell MJ. Suppression of ICE and apoptosis in mammary epithelial cells by extra-cellular matrix. Science. 1995;267:891–3.

    Google Scholar 

  • Chapin RE, Phelps J. Recent advances in testicular cell culture: implications for toxicology. Toxicol In Vitro. 1990;4:543–59.

    Google Scholar 

  • Cheng CY, Mather JP, Byer AL, Bardin CW. Identification of hormonally responsive proteins in primary Sertoli cell culture medium by anion-exchange high performance liquid chromatography. Endocrinology. 1986;118:480–8.

    Google Scholar 

  • Chomczynski P, Sacchi N. Single step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156–9.

    Google Scholar 

  • Clermont Y, Morales CR, Hermo L. Endocytic activities of Sertoli cells in the rat}. Ann NY Acad Sci. 1987;513:1–15.

    Google Scholar 

  • Conzen SD, Cole CN. The three transforming regions of SV40 T antigen are required for immortalization of primary mouse embryo fibroblasts. Oncogene. 1995;11:2295–302.

    Google Scholar 

  • Dadoune J-P, Demoulin A. Structure and functions of the testis. In: Thibault C, Levasseur M-C, Hunter RHF, eds. Reproduction in mammals and man. Paris: Ellipses; 1993:227–55.

    Google Scholar 

  • Daya-Grosjean L, James MR, Drougard C, Sarasin A. An immortalized xeroderma pigmentosum, group C, cell line which replicates SV40 shuttle vectors. Mutat Res. 1987;183:185–96.

    Google Scholar 

  • De Kretser DM, Kerr JB. The cytology of the testis. In: Nobil E et al., eds. The physiology of reproduction. New York: Raven Press; 1988;1:837–932.

    Google Scholar 

  • Frisch SM, Francis H. Disruption of epithelial cell-matrix interactions induces apoptosis. J Cell Biol. 1994;124:619–26.

    Google Scholar 

  • Fritz I, Burdzy K. Novel action of carnitine: inhibition of aggregation of dispersed cells elicited by clusterin in vitro. J Cell Physiol. 1989;140:18–28.

    Google Scholar 

  • Grima J, Pineau C, Bardin CW, Cheng CY. Rat Sertoli cell clusterin, α2-macroglobulin and testins: biosynthesis and differential regulation by germ cells. Mol Cell Endocrinol. 1992;89:127–40.

    Google Scholar 

  • Groome NP. Immunoassays of proteins and anti-peptide anti-bodies. In: Wisdom GB, ed. Peptide antigens. New York: Oxford University Press; 1994:139–79.

    Google Scholar 

  • Guillou F. Characterization of a new superfusion, two-com-partment culture system for Sertoli cells: influence of extra-cellular matrix on the cell permeability and dynamics of transferrin secretion. J Androl. 1990;11:182–94.

    Google Scholar 

  • Guillou F, Zakin MM, Part D, Boissier F, Schaeffer E. Sertoli cell-specific expression of the human transferrin gene. J Biol Chem. 1991;266:9876–84. ai]Hadley MA, Byers SW, Suarez-Quian CA, Kleinman HK, Dym M. Extracellular matrix regulates Sertoli cell differentiation, testicular cord formation, and germ cell development in vitro. J Cell Biol. 1985;101:1511–22.

    Google Scholar 

  • Hansen R, Reddel R, Braithwaite A. The transforming onco-proteins determine the mechanism by which p53 suppresses cell transformation: pRB mediated growth arrest or apoptosis. Oncogene. 1995;11:2535–45.

    Google Scholar 

  • Huang HF, Podach LM, Nathan E, Giglio W. Acute and chronic effects of cisplatinum upon testicular function in rat. J Androl. 1990;11:436–45.

    Google Scholar 

  • Jégou B. The Sertoli cell. Baillière's Clinical Endocrinol Metab. 1992;6:273–331.

    Google Scholar 

  • Jost A, Magre S. Sexual differentiation. In: Thibault C, Levasseur M-C, Hunter RHF, eds. Reproduction in mammals and man. Paris: Ellipses; 1993:197–212.

    Google Scholar 

  • Lash A, Saleem A. Iron metabolism and its regulation. Ann Clin Lab Sci. 1995;25:20–30.

    Google Scholar 

  • Lejeune H, Skalli M, Chatelain PG, Avallet O, Seaz JM. The paracrine role of Sertoli cells on Leydig cell function. Cell Biol Toxicol. 1992;8:73–83.

    Google Scholar 

  • Lewin B. Oncogenes: gene expression and cancer. In: Genes V, ed. Oxford: Oxford University Press and Cell Press; 1994:1181–229.

  • Masson M-T, Bonnet M-C, Lagelle F, Pognan F. Comparative morphology of rat Sertoli cells in testicular sections, in primary cell cultures and in an established cell line (SerW3). Proceedings of the 11th European Congress on Microscopy, Dublin, Ireland; 1996.

  • Matsumoto K, Moriuchi T, Koji T, Nakane PK. Molecular cloning of cDNA coding for rat proliferating cell nuclear antigen (PCNA)/cyclin. EMBO J. 1987;6:637–42.

    Google Scholar 

  • Matter JP. Establishment and characterization of two distinct mouse testicular epithelial cell lines. Biol Reprod. 1980;23:243–52.

    Google Scholar 

  • Morales CR, Clermont Y. Receptor-mediated endocytosis of transferrin by Sertoli cells of the rat. Biol Reprod. 1986;35:393–405.

    Google Scholar 

  • Morales CR, Clermont Y. Phagocytosis and endocytosis in Sertoli cells of the rat. Bull Assoc Anat. 1991;75:157–62.

    Google Scholar 

  • Nambu A, Kumamoto A, Mikuma Y. Effect of anti-cancer agents on cultured rat Sertoli cells. Nipp Hinyokika Gakka " Zasshi. 1995;86:1132–6.

    Google Scholar 

  • Niederberger CS, Shubhada S, Kim SJ, Lamb DJ. Paracrine factors and the regulation of spermatogenesis. World J Urol. 1993;11:120–8.

    Google Scholar 

  • Pineau C, Le Magueresse B, Courtens J-L, Jégou B. Study in vitro of the phagocytic function of Sertoli cells in the rat. Cell Tissue Res. 1991;264:589–98.

    Google Scholar 

  • Pineau C, Syed V, Bardin CW, Jégou B, Cheng CY. Germ cell-conditioned medium contains multiple factors that modulate the secretion of testins, clusterin and transferrin by Sertoli cells. J Androl. 1993;14:87–98.

    Google Scholar 

  • Rosenberg NH, Yuan LA, Reyaz QX, Bhasin S. Transcriptional regulation of inhibin b messenger ribonucleic acid levels in TM4 or primary rat Sertoli cells by 8-bromo-cyclic adeno-sine monophosphate. Mol Endocrinol. 1993;7:561–9.

    Google Scholar 

  • Russel LD, Ettlin RA, Sinha-Hikim AP, Clegg ED. Mammalian spermatogenesis. In: Histological and histopathological evaluation of the testis. Clearwater: Cache River Press; 1990:1–40.

  • Sambrook J, Fritsch EF, Maniatis T. Molecular cloning, a laboratory manual. 2nd ed. Cold Spring Harbor: Cold Spring Harbor Laboratory Press; 1989.

  • Sherr CJ. G1 phase progression: cycling on cue. Cell. 1994;79:551–5.

    Google Scholar 

  • Shubhada S, Glinz M, Lamb DJ. Sertoli cell secreted growth factor: cellular origin, paracrine and endocrine regulation of secretion. J Androl. 1993;14:99–109.

    Google Scholar 

  • Symonds H, Krall L, Remington L et al. p53 dependent apoptosis suppresses tumor growth and progression in vivo. Cell. 1994;78:703–11.

    Google Scholar 

  • Tiemann F, Zerrahn J, Deppert W. Cooperation of Simian virus 40 large and small T antigens in metabolic stabilization of tumor suppressor p53 during cellular transformation. J Virol. 1995;69:6115–21.

    Google Scholar 

  • Verhoeven G, Cailleau J. Testicular peritubular cells secrete a protein under androgen control that inhibits induction of aromatase activity in Sertoli cells. Endocrinology. 1988;123:2100–10.

    Google Scholar 

  • Wauben-Penris PJJ, Strous GJ, Van der Donk HA. Kinetics of transferrin endocytosis and iron uptake by intact isolated rat seminiferous tubules and Sertoli cells in culture. Biol Reprod. 1988;38:853–61.

    Google Scholar 

  • Xi DR, Djakiew D, Dym M. Endocytic activity of Sertoli cells grown in bicameral culture chambers. Anat Rec. 1987;218:306–12.

    Google Scholar 

  • Xiong Y, Zhang H, Beach D. Subunit rearrangement of the cyclin-dependent kinases is associated with cellular transformation. Genes Dev. 1993;7:1572–83.

    Google Scholar 

  • Yonish-Rouach E, Resnitzky D, Lotem J, Sachs L, Kimchi A, Oren M. Wild-type p53 induces apoptosis of myeloid leukaemic cells that is inhibited by interleukin-6. Nature. 1991;352:345–7.

    Google Scholar 

  • Zwain I, Grima J, Stahler S et al. Regulation of Sertoli cell α2-macroglobulin and clusterin (SGP2) secretion by peritubu-lar myoid cells. Biol Reprod. 1993;48:180–7.

    Google Scholar 

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Authors and Affiliations

  1. Drug Safety Evaluation, Pfizer Central Research, Amboise, France

    F. Pognan, M.-T. Masson, F. Lagelle & C. Charuel

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  1. F. Pognan
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  2. M.-T. Masson
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  3. F. Lagelle
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  4. C. Charuel
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Pognan, F., Masson, MT., Lagelle, F. et al. Establishment of a rat Sertoli cell line that displays the morphological and some of the functional characteristics of the native cell. Cell Biol Toxicol 13, 453–463 (1997). https://doi.org/10.1023/A:1007475928452

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