Human Female Reproductive Tract Epithelial Cell Culture

The female reproductive system is a complex system. Epithelia of the female reproductive system including the ovaries, the oviduct, and the uterus are important sites for follicular development, ovulation, fertilization, implantation, and embryo development. They are also able to synthesize and secrete various hormones, growth factors, and cytokines, which are essential to women’s health, sexuality, and reproduction. Conversely, their dysfunction has been implicated in disorders such as infertility, endometriosis, and many other gynecological diseases, as well as cancer. In this chapter, we describe detailed procedures for establishing and maintaining primary cultures of human ovarian surface epithelium, oviductal epithelium, and endometrium. We also provide protocols for cell immortalization, clonal isolation, and in coculture with stromal cells. These cultures can be useful models for investigating the molecular and cellular functions of these epithelia in both normal and pathological states. Access this chapter Tax calculation will be finalised at checkout Purchases are for personal use only Similar content being viewed by others

Auersperg N, Wong AS, Choi KC, Kang SK, Leung PC (2001) Ovarian surface epithelium: biology, endocrinology, and pathology. Endocr Rev 22:255–288 Kruk PA, Maines-Bandiera SL, Auersperg N (1990) A simplified method to culture human ovarian surface epithelium. Lab Invest 63: 132–136 Maines-Bandiera SL, Kruk PA, Auersperg N (1992) Simian virus 40-transformed human ovarian surface epithelial cells escape normal growth controls but retain morphogenetic responses to extracellular matrix. Am J Obstet Gynecol 167:729–735 Tsao SW, Mok SC, Fey EG, Fletcher JA, Wan TS, Chew EC, Muto MG, Knapp RC, Berkowitz RS (1995) Characterization of human ovarian surface epithelial cells immortalized by human papilloma viral oncogenes (HPV-E6E7 ORFs). Exp Cell Res 218: 499–507 Kohler MF, Kerns BJ, Humphrey PA, Marks JR, Bast RC Jr, Berchuck A (1993) Mutation and overexpression of p53 in early-stage epithelial ovarian cancer. Obstet Gynecol 81: 643–650 Oishi T, Kigawa J, Minagawa Y, Shimada M, Takahashi M, Terakawa N (1998) Alteration of telomerase activity associated with development and extension of epithelial ovarian cancer. Obstet Gynecol 91:568–571 Baykal A, Thompson JA, Xu XC, Hahn WC, Deavers MT, Malpica A, Gershenson DM, Silva EG, Liu J (2004) In situ human telomerase reverse transcriptase expression pattern in normal and neoplastic ovarian tissues. Oncol Rep 11:297–302 Yao YQ, Ho PC, Yeung WSB (1999) Effects of human oviductal cell coculture on various functional parameters of human spermatozoa. Fertil Steril 71:232–239 Yeung WSB, Xu JS, Lee CKF (2002) The oviduct and preimplantation embryo development. Reprod Med Rev 10:21–44 Lee YL, Lee KF, Xu JS, He QY, Chiu JF, Lee WM, Luk JM, Yeung WSB (2004) The embryotrophic activity of oviductal cell-derived complement C3b and iC3b, a novel function of complement protein in reproduction. J Biol Chem 279:12763–12768 Lee KF, Xu JS, Lee YL, Yeung WSB (2006) Demilune cell and parotid protein from murine oviductal epithelium stimulates preimplantation embryo development. Endocrinology 147:79–87 Levanon K, Crum C, Drapkin R (2008) New insights into the pathogenesis of serous ovarian cancer and its clinical impact. J Clin Oncol 26:5284–5293 Seppälä M, Taylor RN, Koistinen H, Koistinen R, Milgrom E (2002) Glycodelin: a major lipocalin protein of the reproductive axis with diverse actions in cell recognition and differentiation. Endocr Rev 23:401–430 Horne AW, Lalani EN, Margara RA, White JO (2006) The effects of sex steroid hormones and interleukin-1-beta on MUC1 expression in endometrial epithelial cell lines. Reproduction 131:733–742 Robertson SA (2007) GM-CSF regulation of embryo development and pregnancy. Cytokine Growth Factor Rev 18:287–298 Domínguez F, Gadea B, Esteban FJ, Horcajadas JA, Pellicer A, Simón C (2008) Comparative protein-profile analysis of implanted versus non-implanted human blastocysts. Hum Reprod 23:1993–2000 Dharmaraj N, Gendler SJ, Carson DD (2009) Expression of human MUC1 during early pregnancy in the human MUC1 transgenic mouse model. Biol Reprod 81:1182–1188 Lam KK, Chiu PC, Chung MK, Lee CL, Lee KF, Koistinen R, Koistinen H, Seppala M, Ho PC, Yeung WSB (2009) Glycodelin-A as a modulator of trophoblast invasion. Hum Reprod 24:2093–2103 Lee CL, Chiu PC, Lam KK, Chan RW, Chu IK, Koistinen R, Koistinen H, Seppälä M, Lee KF, Yeung WSB (2010) Glycodelin-A modulates cytokine production of peripheral blood natural killer cells. Fertil Steril 94:769–771 Gargett CE, Schwab KE, Zillwood RM, Nguyen HP, Wu D (2009) Isolation and culture of epithelial progenitors and mesenchymal stem cells from human endometrium. Biol Reprod 80:1136–1145 Gargett CE, Chan RW, Schwab KE (2007) Endometrial stem cells. Curr Opin Obstet Gynecol 19:377–383 Chatot CL, Ziomek CA, Bavister BD, Lewis JL, Torres I (1989) An improved culture medium supports development of random-bred 1-cell mouse embryos in vitro. J Reprod Fertil 86:679–688 Auersperg N, Siemens CH, Myrdal SE (1984) Human ovarian surface epithelium in primary culture. In Vitro 20:743–755 Wong AS, Maines-Bandiera SL, Leung PC, Auersperg N (1998) Metaplastic changes in cultured ovarian surface epithelium. In Vitro Cell Dev Biol 34:668–670 Wong AS, Maines-Bandiera SL, Rosen B, Wheelock MJ, Johnson KR, Leung PC, Roskelley CD, Auersperg N (1999) Constitutive and conditional cadherin expression in cultured human ovarian surface epithelium: influence of family history of ovarian cancer. Int J Cancer 81:180–188 Wong AS, Pelech SL, Woo MM, Yim G, Rosen B, Ehlen T, Leung PC, Auersperg N (2001) Coexpression of hepatocyte growth factor-Met: as an early step in ovarian carcinogenesis? Oncogene 20:1318–1328 Auersperg N, Maines-Bandiera SL, Dyck HG, Kruk PA (1994) Characterization of cultured human ovarian surface epithelial cells: phenotypic plasticity and premalignant changes. Lab Invest 71:510–518 Fromm M, Berg P (1982) Deletion mapping of DNA regions required for SV40 early region promoter function in vivo. J Mol Appl Genet 1:457–481 Yang G, Rosen DG, Colacino JA, Mercado-Uribe I, Liu J (2007) Disruption of the retinoblastoma pathway by small interfering RNA and ectopic expression of the catalytic subunit of telomerase lead to immortalization of human ovarian surface epithelial cells. Oncogene 26:1492–1498 Horikawa I, Cable PL, Afshari C, Barrett JC (1999) Cloning and characterization of the promoter region of human telomerase reverse transcriptase gene. Cancer Res 59: 826–830 Yang G, Rosen DG, Mercado-Uribe I, Colacino JA, Mills GB, Bast RC Jr, Zhou C, Liu J (2007) Knockdown of p53 combined with expression of the catalytic subunit of telomerase is sufficient to immortalize primary human ovarian surface epithelial cells. Carcinogenesis 28:174–182 Chan RW, Schwab KE, Gargett CE (2004) Clonogenicity of human endometrial epithelial and stromal cells. Biol Reprod 70:1738–1750 Salamanca CM, Maines-Bandiera SL, Leung PC, Hu YL, Auersperg N (2004) Effects of epidermal growth factor/hydrocortisone on the growth and differentiation of human ovarian surface epithelium. J Soc Gynecol Investig 11:241–251 Bongso A, Ng SC, Sathananthan H, Ng PL, Rauff M, Ratnam SS (1989) Establishment of human ampullary cell cultures. Hum Reprod 4:486–494 Lee YL, Lee KF, Xu JS, Wang YL, Tsao SW, Yeung WSB (2001) Establishment and characterization of an immortalized human oviductal cell line. Mol Reprod Dev 59:400–409 Lalitkumar PG, Lalitkumar S, Meng CX, Stavreus-Evers A, Hambiliki F, Bentin-Ley U, Gemzell-Danielsson K (2007) Mifepristone, but not levonorgestrel, inhibits human blastocyst attachment to an in vitro endometrial three-dimensional cell culture model. Hum Reprod 22:3031–3037 Ho Y, Wigglesworth K, Eppig JJ, Schultz RM (1995) Preimplantation development of mouse embryos in KSOM: augmentation by amino acids and analysis of gene expression. Mol Reprod Dev 41:232–238 Pellegrini G, Golisano O, Paterna P, Lambiase A, Bonini S, Rama P, De Luca M (1999) Location and clonal analysis of stem cells and their differentiated progeny in the human ocular surface. J Cell Biol 145:769–782 Acknowledgments We thank the Department of Obstetrics and Gynecology, University of Hong Kong, which kindly provided OSE, oviductal, and endometrial samples. This work was supported by grants from the Hong Kong Research Grant Council to A.S.T.W. and (HKU 782209M) W.S.B.Y. and from the HKU Outstanding Young Researcher Award to A.S.T.W. Author information Authors and Affiliations Corresponding author Editor information Editors and Affiliations Rights and permissions Copyright information © 2012 Springer Science+Business Media, LLC About this protocol Cite this protocol Chan, R.W.S. et al. (2012). Human Female Reproductive Tract Epithelial Cell Culture. In: Randell, S., Fulcher, M. (eds) Epithelial Cell Culture Protocols. Methods in Molecular Biology, vol 945. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-125-7_21 Download citation DOI: https://doi.org/10.1007/978-1-62703-125-7_21 Published: Publisher Name: Humana Press, Totowa, NJ Print ISBN: 978-1-62703-124-0 Online ISBN: 978-1-62703-125-7 eBook Packages: Springer Protocols