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Primary Culture of Ovarian Cells for Research on Cell Interactions in the Hormonal Control of Steroidogenesis

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Human Cell Culture Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 806))

Abstract

Ovary is a polymorphic complex structure in which the cells are arranged in two essential endocrine mini glands: the follicle (F) and the corpus luteum (CL). Their secretory function creates an optimal milieu for growth, maturation, and selection of follicles and oocytes competent for ovulation. Monoculture of isolated ovarian cells has identified the secretory potential of the different cell types functioning in this complex gland in vivo. Primary culture of isolated ovarian cells is a good tool for the investigation of cell interactions and its impact on steroidogenesis, dynamics of steroidogenic enzymes, hormone receptors, changes in the cytoskeleton in granulosa cell populations, regulatory mechanisms, and the intracellular pathways of gonadotropin signaling in steroidogenic ovarian cells. Since the granulosa cell number recovered during isolation from the follicle is substantial, this makes primary culture fairly easy and enables many kinds of studies in vitro. Ovarian cells are highly differentiated and express characteristic functional specificity dependent on the dynamics of the sexual cycle. This is important so as not to produce artifacts in vitro.

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References

  1. Galas, J., Slomczynska, M., Pierściński, A, (2003) Effect of photoperiod on distribution patterns of androgen receptors and steroid hormone concentrations in ovaries of bank voles. Acta histochem. 105, 175–181.

    Article  PubMed  CAS  Google Scholar 

  2. Galas, J., Knapczyk, K., Slomczynska, M., Kwaśnik, A. (2007) Distribution of androgen receptor and steroid hormone concentrations in ovaries of immature bank voles: Effect of photoperiod. Acta histochem. 109, 437–445.

    Article  PubMed  CAS  Google Scholar 

  3. Galas, J., Clarke J.R., Soltys, Z., Stoklosowa,S. (2005) Effect of photoperiod on cultured granulosa cell of the bank vole Cletrionomys glareolus. Comp. Biochem. Physiol. 140(2), 187–194.

    Google Scholar 

  4. Stoklosowa, S., Gregoraszczuk, E.L., Channing, C.P. (1982) Estrogen and progesterone secretion by isolated cultured porcine thecal and granulosa cells. Biol. Reprod. 26, 943–952.

    Article  PubMed  CAS  Google Scholar 

  5. Lei, Z.M., Chegini, N., Rao, C.H. (1991) Quantitative cell composition of human and bovine corpora lutea from various reproductive states. Reproduction, 44, 1148–1156.

    Article  CAS  Google Scholar 

  6. Gregoraszczuk, E. L. (1992) Interrelationship in steroid hormone secretion and morphological changes of porcine corpora lutea at various periods of luteal phase. Endocrine Regulations. 26, 189–194.

    PubMed  CAS  Google Scholar 

  7. Fitz, T.A., Mayan,M.H., Sawyer H.R., Niswender, G.D. (1982) Characteris of two steroidogenic cell types in the ovine corpus luteum. Biol. Reprod., 27, 703–711.

    Article  PubMed  CAS  Google Scholar 

  8. Fish,B., Margara, R.A., Winston, R.M.L., Hillier, S.G. (1989) Cellular basis of luteal steroidogenesis in the human ovary. J. Endocrinol., 122, 303–311.

    Article  Google Scholar 

  9. Mass-Szabowski N., Stark H.J., Fusenig N.E. (2000) Keratinocite growth regulation in defined organotypic cultures through Il-1 induced keratinocyte growth factor expression in resting fibroblasts. J. Invest. Dermatol. 114, 1075–1084.

    Article  Google Scholar 

  10. Channing, C.P., Ledwitz–Righby, F. Methods for assessing hormone mediated differentiation of ovarian cells in culture and short term incubation. In: Colowick, S.P., Kaplan, N.O, Methods in Enzymology, XXXIX, Part D., Hartman, J.B., O’Malley, B., (eds), Academic Press, New York, London, (1975).

    Google Scholar 

  11. Karamouti, M., Kaltitsaris, A, Vamvakopaulos, N., Kotlios, G., Messinis J.E. (2008) Growth hormone, insulin-like growth factor I, and leptin interaction in human cultured lutein granuloza cell steroidogensis. Fertility Sterility 90, 1444–50.

    Article  CAS  Google Scholar 

  12. Fingenschau, Y.,Sandsfjord, J.A., Yousef, M.I., Fuskevag, O.M., Sveinbjornson, B., Bertheussen, K. (1997) A simplified serum-free method for preparation and cultivation of human granulose luteal cells. Human Reproduction 12, 523–531.

    Article  Google Scholar 

  13. Taketani, T., Yamagata, Y., Takasaki, A., Matsuoka, A., Tamura, H., Sugino, N. (2008) Effects of growth hormone and insulin-like growth factor-1 on progesterone production in human luteinized granulosa cells. Fertility and Sterility 90, 744–48.

    Article  PubMed  CAS  Google Scholar 

  14. Stoklosowa, S. (1982) Tissue culture of gonadal cells. Acta biol. Acad. Sci.hung. 33(4), 367–379.

    Google Scholar 

  15. Alila, Rogo, K. O., Gombe S. (1987) Effects of prolactin on steroidogenesis by human luteal cells in culture. Fertility and Sterility 47, 947–955.

    Google Scholar 

  16. Stoklosowa, S. (2001) Three dimentional tissue and organ models in vitro: their application in basic and practical research. Folia Histochem. Cytobiol. 39, 91–96.

    PubMed  CAS  Google Scholar 

  17. Gregoraszczuk, E.L. (1995) Effect of FSH on progesterone secretion by porcine large and small luteal cells isolated from early, developing corpora lutea. Exp. Clin. Endocrinol. 103, 272–274.

    Article  CAS  Google Scholar 

  18. Cell and tissue culture (in Polish): Hodowla komórek i tkanek S. Stoklosowa (ed) PWN, Warszawa, 2006

    Google Scholar 

  19. Kmicikiewicz, I., Wojtusiak, A., Bilinska, B. (1999) The effect of testicular macrophages, macrophages-conditioned medium and interleukin-1 on bank vole Leydig cell steroidogenesis. Exptl. Clin. Endocrinol. Diabetes. 107, 262–271.

    Article  CAS  Google Scholar 

  20. Freshney, R.I. Culture of Animal Cells. Wiley-Liss N. Y (2004).

    Google Scholar 

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Acknowledgments

Advice of Professor S. Stoklosowa from the Department of Endocrinology, Jagiellonian University was greatly appreciated.

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Correspondence to Jerzy F. Galas .

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Galas, J.F. (2012). Primary Culture of Ovarian Cells for Research on Cell Interactions in the Hormonal Control of Steroidogenesis. In: Mitry, R., Hughes, R. (eds) Human Cell Culture Protocols. Methods in Molecular Biology, vol 806. Humana Press. https://doi.org/10.1007/978-1-61779-367-7_16

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  • DOI: https://doi.org/10.1007/978-1-61779-367-7_16

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-366-0

  • Online ISBN: 978-1-61779-367-7

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