{"paper_id":"1e166071-45ec-4ac8-bd30-1a95464eb2c6","body_text":"Abstract\nThe 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.\nAccess this chapter\nTax calculation will be finalised at checkout\nPurchases are for personal use only\nSimilar content being viewed by others\nReferences\nAuersperg N, Wong AS, Choi KC, Kang SK, Leung PC (2001) Ovarian surface epithelium: biology, endocrinology, and pathology. Endocr Rev 22:255–288\nKruk PA, Maines-Bandiera SL, Auersperg N (1990) A simplified method to culture human ovarian surface epithelium. Lab Invest 63: 132–136\nMaines-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\nTsao 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\nKohler 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\nOishi 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\nBaykal 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\nYao YQ, Ho PC, Yeung WSB (1999) Effects of human oviductal cell coculture on various functional parameters of human spermatozoa. Fertil Steril 71:232–239\nYeung WSB, Xu JS, Lee CKF (2002) The oviduct and preimplantation embryo development. Reprod Med Rev 10:21–44\nLee 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\nLee 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\nLevanon 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\nSeppälä 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\nHorne 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\nRobertson SA (2007) GM-CSF regulation of embryo development and pregnancy. Cytokine Growth Factor Rev 18:287–298\nDomínguez 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\nDharmaraj 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\nLam 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\nLee 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\nGargett 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\nGargett CE, Chan RW, Schwab KE (2007) Endometrial stem cells. Curr Opin Obstet Gynecol 19:377–383\nChatot 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\nAuersperg N, Siemens CH, Myrdal SE (1984) Human ovarian surface epithelium in primary culture. In Vitro 20:743–755\nWong AS, Maines-Bandiera SL, Leung PC, Auersperg N (1998) Metaplastic changes in cultured ovarian surface epithelium. In Vitro Cell Dev Biol 34:668–670\nWong 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\nWong 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\nAuersperg 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\nFromm 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\nYang 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\nHorikawa 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\nYang 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\nChan RW, Schwab KE, Gargett CE (2004) Clonogenicity of human endometrial epithelial and stromal cells. Biol Reprod 70:1738–1750\nSalamanca 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\nBongso A, Ng SC, Sathananthan H, Ng PL, Rauff M, Ratnam SS (1989) Establishment of human ampullary cell cultures. Hum Reprod 4:486–494\nLee 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\nLalitkumar 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\nHo 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\nPellegrini 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\nAcknowledgments\nWe 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.\nAuthor information\nAuthors and Affiliations\nCorresponding author\nEditor information\nEditors and Affiliations\nRights and permissions\nCopyright information\n© 2012 Springer Science+Business Media, LLC\nAbout this protocol\nCite this protocol\nChan, 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\nDownload citation\nDOI: https://doi.org/10.1007/978-1-62703-125-7_21\nPublished:\nPublisher Name: Humana Press, Totowa, NJ\nPrint ISBN: 978-1-62703-124-0\nOnline ISBN: 978-1-62703-125-7\neBook Packages: Springer Protocols","source_license":"CC0","license_restricted":false}