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Isolation and characterization of rabbit ovarian surface epithelium, granulosa cells, and peritoneal mesothelium in primary culture

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Summary

Mammalian ovarian surface epithelial (OSE) cells and peritoneal mesothelial (PM) cells have a common embryologic origin, yet certain morphologic and histochemical characteristics are different in the adult. In this study, a two-step culture method was developed to examine the characteristics of these two cell types in vitro. OSE, PM, and ovarian granulosa (GC) cells were isolated from estrous rabbits and cultured for 6 d in 5% serum-supplementedd-valine medium (to inhibit fibroblast growth), then incubated for a further 2 d in serum-free McCoy's 5A medium. This study showed that rabbit OSE and PM cells in vitro maintained certain in vivo morphologic characteristics; OSE cells exhibited distinct cell borders and abundant microvilli of homogeneous size and shape, whereas PM cells were characterized by obscure cell borders and abundant microvilli of heterogeneous form. GC in vitro exhibited overlapping cell borders and sparse microvilli of homogeneous structure. This study showed for the first time that cultured rabbit OSE and PM cells, but not GC, contain distinct filaments of cytokeratin 18. In addition, rabbit OSE cells and GC, but not PM cells, contained 17β-hydroxysteroid dehydrogenase. However, only GC contained delta 5-3β hydroxysteroid dehydrogenase. OSE, PM, and GC maintained their ultrastructural and histochemical characteristics in serum-free medium. These results suggest that rabbit OSE cells in vitro could be distinguished from PM cells by histochemical and ultrastructural differences. Furthermore, because these characteristics were not altered in serum-free medium, the two-step culture method will be valuable in further hormonal studies of these cells in vitro.

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References

  1. Scully, R. E. Ovarian tumors—a review. Am. J. Pathol. 87:686–720; 1977.

    PubMed  CAS  Google Scholar 

  2. Falkson, C. I. Mesothelioma or ovarian cancer? S. Afr. Med. J. 68:676–677; 1985.

    PubMed  CAS  Google Scholar 

  3. Parmley, T. H.; Woodruff, J. D. The ovarian mesothelioma. Am. J. Obstet. Gynecol. 120:234–241; 1974.

    PubMed  CAS  Google Scholar 

  4. Foyle, A.; Al-Jabi, M.; McCaughey, W. T. E. Papillary peritoneal tumors in women. Am. J. Surg. Pathol. 5:241–249, 1981.

    Article  PubMed  CAS  Google Scholar 

  5. Genadry, R.; Poliakoff, S.; Rotmensch, J., et al. Primary, papillary peritoneal neoplasia. Obstet. Gynecol. 58:730–734; 1981.

    PubMed  CAS  Google Scholar 

  6. Swerdlow, M. Mesothelioma of the pelvic peritoneum resembling papillary cystadenocarcinoma of ovary. Am. J. Obstet. Gynecol. 77:197–200; 1959.

    PubMed  CAS  Google Scholar 

  7. Julian, C. G.; Goss, J.; Blanchard K., et al. Biological behaviour of primary ovarian malignancy. Obstet. Gynecol. 44:873–884; 1974.

    PubMed  CAS  Google Scholar 

  8. Blaustein, A. Peritoneal mesothelium and ovarian surface cells-shared characteristics. Int. J. Gynecol. Pathol. 3:361–375; 1984.

    Article  PubMed  CAS  Google Scholar 

  9. Blaustein, A. U.; Lee, H. Surface cells of the ovary and pelvic peritoneum. A histochemical and ultrastructural comparison. Gynecol. Oncol. 8:34–43; 1979.

    Article  PubMed  CAS  Google Scholar 

  10. Hoang-Noc, M.; Smadja, A.; Orcel, L. Etude morphologique comparée du mésothélium péritonéal et de l'épithélium germinatif de l'ovaire. J. Gynecol. Obstet. Biol. Reprod. 17:479–484; 1988.

    Google Scholar 

  11. Nicosia, S. V.; Johnson, J. H. Surface morphology of ovarian mesothelium (surface epithelium) and of other pelvic and extrapelvic mesothelial sites in the rabbit. Int. J. Gynecol. Pathol. 3:249–260; 1984.

    Article  PubMed  CAS  Google Scholar 

  12. Klein, B.; Falkson, G. Ovarian cancer. S. Afr. Cancer. Bull. 27:140–145; 1983.

    Google Scholar 

  13. Lerner, H. J.; Schoenfeld, D. A.; Martin, A., et al. Malignant mesothelioma: ECOG experience. Cancer 52:1981–1985; 1983.

    Article  PubMed  CAS  Google Scholar 

  14. Kannerstein, M.; Churg, J.; McCaughey, W. T. E., et al. Papillary tumors of the peritoneum in women; mesothelioma or papillary carcinoma. Am. J. Obstet. Gynecol. 127:306–314; 1977.

    PubMed  CAS  Google Scholar 

  15. Hamilton, T. C.; henderson, W. J.; Eaton, C. Isolation and growth of the rat ovarian germinal epithelium. In: Richards, R. J.; Rajan, K. T., eds. Tissue culture in medical research, II: proceedings of 2nd international symposium. New York: Pergamon Press. 1980:237–244.

    Google Scholar 

  16. Adams, A. T.; Auersperg, N. Transformation of cultured rat ovarian surface epithelium cells by kirsten murine sarcoma virus. Cancer Res 41:2063–2072; 1981.

    PubMed  CAS  Google Scholar 

  17. Auersperg, N.; Siemens, C. H.; Myrdal, S. E. Human ovarian surface epithelium in primary culture. In Vitro 20:743–755; 1984.

    Article  PubMed  CAS  Google Scholar 

  18. Nicosia, S. V.; Johnson, J. H.; Streibel, E. J. Isolation and ultrastructure of rabbit ovarian mesothelium (surface epithelium). Int. J. Gynecol. Pathol. 3:348–360; 1984.

    Article  PubMed  CAS  Google Scholar 

  19. Nicholson, L. J.; Clark, J. M. F.; Pittilo R. M., et al. The mesothelial cell as a nonthrombogenic surface. Thromb. Haemostasis. 52:2511–2514; 1984.

    Google Scholar 

  20. Gilbert, S. F.; Migeon, B. R.d-Valine as a selective agent for normal human and rodent epithelial cells in culture. Cell 5:11–17; 1975.

    Article  PubMed  CAS  Google Scholar 

  21. Spurr, A. R. A low-viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res. 26:31–43; 1969.

    Article  PubMed  CAS  Google Scholar 

  22. Gamliel, H. Optimum fixation conditions may allow air drying of soft biological specimens with minimum cell shrinkage and maximum preservation of surface features. Scan. Electron. Microsc. (Pt. 4):1649–1664; 1985.

  23. Tölle, H.-G.; Weber, K.; Osborn, M., Microinjection of monoclonal antibodies specific for one intermediate filament protein in cells containing multiple keratins allow insight into the composition of particular 10 nm filaments. Eur. J. Cell Biol. 38:234–244; 1985.

    PubMed  Google Scholar 

  24. Levy, H.; Deane, H. W.; Rubin, B. L. Visualization of steroid-3β-Ol-dehydrogenase activity in tissue of intact and hypophysectomized rats. Endocrinology 65:932–943; 1959.

    Article  PubMed  CAS  Google Scholar 

  25. Anderson, E.; Selig, M.; Lee, G., et al. Androgen-induced changes in ovarian granulosa cells from immature ratsin vitro. In: Mahesh, V. B.; Dhindsa, D. S.; Anderson, E. et al. eds. Regulation of ovarian and testicular function. New York: Plenum Press. 1987:259–274.

    Google Scholar 

  26. Stein, K.; Allen, E. Attempts to stimulate proliferation of the germinal epithelium of the ovary. Anat. Rec. 82:1–9; 1942.

    Article  Google Scholar 

  27. Hamilton, T. C.; Davies, P.; Griffiths, K. Androgen and estrogen binding in cytosols of human ovarian tumours. J. Endocrinol. 90:421–431; 1981.

    Article  PubMed  CAS  Google Scholar 

  28. Hamilton, T. C.; Davies, P.; Griffiths, K. Oestrogen receptor-like binding in the surface germinal epithelium of the rat ovary. J. Endocrinol. 95:377–385; 1982.

    Article  PubMed  CAS  Google Scholar 

  29. Adams, A. T.; Auersperg, N. Autoradiographic investigation of estrogen binding in cultured rat ovarian surface epithelial cells. J. Histochem. Cytochem. 31:1321–1325; 1983.

    PubMed  CAS  Google Scholar 

  30. Simon, W. E.; Albrecht, M.; Hansel, M. et al. Cell lines derived from human ovarian carcinomas: growth stimulation by gonadotropin and steroid hormones. JNCI 70:839–844; 1983.

    PubMed  CAS  Google Scholar 

  31. Osterholzer, H. O.; Streibel, E. J.; Nicosia, S. V. Growth effects of protein hormones on cultured rabbit ovarian surface epithelial cells. Biol. Reprod. 33:247–258; 1985.

    Article  PubMed  CAS  Google Scholar 

  32. Rajaniemi, H.; Kauppila, A.; Ronnberg, L., et al. LH (hCG) receptor in benign and malignant tumors of human ovary. Acta Obstet. Gynecol. Scand. Suppl. 101:83–86; 1981.

    Article  PubMed  CAS  Google Scholar 

  33. Cramer, D. W.; Hutchison, G. B.; Welch, W. R., et al. Factors affecting the association of oral contraceptives and ovarian cancer. N. Engl. J. Med. 307:1047–1051; 1982.

    Article  PubMed  CAS  Google Scholar 

  34. Resta, L.; De Benedictis, G.; Scordari, M. D., et al. Hyperplasia and metaplasia of ovarian surface epithelium in women with endometrial carcinoma. Suggestion for a hormonal influence in ovarian carcinogenesis. Tumori 73:249–256; 1987.

    PubMed  CAS  Google Scholar 

  35. Schwartz, N. B. The role of FSH and LH and of their antibodies on follicle growth and on ovulation. Biol. Reprod. 10:236–272; 1974.

    Article  PubMed  CAS  Google Scholar 

  36. Channing, C. P.; Tsafriri, A. Mechanism of action of luteinizing hormone and follicle-stimulating hormone on the ovaryin vitro. Metabolism 26:413–468; 1977.

    Article  PubMed  CAS  Google Scholar 

  37. Fathalla, M. F. Incessant ovulation—a factor in ovarian neoplasm? Lancet 2:163; 1971.

    Article  PubMed  CAS  Google Scholar 

  38. Zajicek, J. Ovarian cystomas and ovulations, a histogenetic concept. Tumori 63:429–435; 1977.

    PubMed  CAS  Google Scholar 

  39. Osterholzer, H. O.; Johnson, J. E.; Nicosia, S. V. An autoradiographic study of rabbit ovarian surface epithelium before and after ovulation. Biol. Reprod. 33:729–738; 1985.

    Article  PubMed  CAS  Google Scholar 

  40. Siemens, C.; Auersperg N. Serial propagation of human ovarian surface epithelium in tissue culture. J. Cell. Physiol. 134:347–356; 1988.

    Article  PubMed  CAS  Google Scholar 

  41. Piquette, G. N.; Timms, B. G. Immunohistochemical localization of tissue-type plasminogen activator (t-PA) in primary cultures of rabbit ovarian surface epithelium, granulosa cells, and peritoneal mesothelium. Biol. Reprod. 38(Suppl. 1:Abs) 405; 1988.

    Google Scholar 

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Authors and Affiliations

  1. School of Medicine, Department of Anatomy, University of South Dakota, 57069, Vermillion, South Dakota

    Gary N. Piquette & Barry G. timms

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  1. Gary N. Piquette
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  2. Barry G. timms
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This work was supported in part by Grant No. 202-3192 from the University of South Dakota Parsons Fund

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Piquette, G.N., timms, B.G. Isolation and characterization of rabbit ovarian surface epithelium, granulosa cells, and peritoneal mesothelium in primary culture. In Vitro Cell Dev Biol 26, 471–481 (1990). https://doi.org/10.1007/BF02624089

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  • DOI: https://doi.org/10.1007/BF02624089

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