Abstract
We have refined the technique for isolating and propagating cultures of primary epithelial ovarian cancer (EOC) cells derived from solid tumors and ascites. Both protocols involve a simple yet rapid method for the growth and propagation of EOC tumor and ascites cells in a basal culture medium without the addition of growth factors. Isolation of tumor EOC cells involves the mechanical disruption of the tumor tissue with the help of a cell scraper, while ascites-derived EOC cells are mixed with growth medium and placed directly into culture with very little manipulation. We further describe a partial trypsinization method to eliminate fibroblast contamination from primary EOC cells derived from solid tumors. These methods allow for the direct application of many molecular, cellular, and functional analyses within a few weeks of initial isolation, with the added potential of retrospective analyses of archived cells and tissues. Thus, we have included steps for long-term cryopreservation of early-passage EOC cells. Initial isolation of EOC cells can be completed within 1 h, and primary cells are further expanded in culture for several weeks.
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References
Siegel R, Ward E, Brawley O, Jemal A (2011) Cancer statistics, 2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA Cancer J Clin 61:212–236
Weberpals JI, Koti M, Squire JA (2011) Targeting genetic and epigenetic alterations in the treatment of serous ovarian cancer. Cancer Genet 204:525–535
Saad AF, Hu W, Sood AK (2010) Microenvironment and pathogenesis of epithelial ovarian cancer. Horm Cancer 1:277–290
Asadollahi R, Hyde CA, Zhong XY (2010) Epigenetics of ovarian cancer: from the lab to the clinic. Gynecol Oncol 118:81–87
Yoshida S, Furukawa N, Haruta S, Tanase Y, Kanayama S, Noguchi T, Sakata M, Yamada Y, Oi H, Kobayashi H (2009) Expression profiles of genes involved in poor prognosis of epithelial ovarian carcinoma: a review. Int J Gynecol Cancer 19:992–997
Sheng Q, Liu J (2011) The therapeutic potential of targeting the EGFR family in epithelial ovarian cancer. Br J Cancer 104:1241–1245
Eckstein N (2011) Platinum resistance in breast and ovarian cancer cell lines. J Exp ClinCancer 30:91
van Jaarsveld MT, Helleman J, Berns EM, Wiemer EA (2010) MicroRNAs in ovarian cancer biology and therapy resistance. Int J Biochem Cell Biol 42:1282–1290
Grun B, Benjamin E, Sinclair J, Timms JF, Jacobs IJ, Gayther SA, Dafou D (2009) Three-dimensional in vitro cell biology models of ovarian and endometrial cancer. Cell Prolif 42:219–228
Theriault BL, Shepherd TG, Mujoomdar ML, Nachtigal MW (2007) BMP4 induces EMT and Rho GTPase activation in human ovarian cancer cells. Carcinogenesis 28:1153–1162
Matsumura N, Huang Z, Mori S, Baba T, Fujii S, Konishi I, Iversen ES, Berchuck A, Murphy SK (2011) Epigenetic suppression of the TGF-beta pathway revealed by transcriptome profiling in ovarian cancer. Genome Res 21:74–82
Ouellet V, Provencher DM, Maugard CM, Le Page C, Ren F, Lussier C, Novak J, Ge B, Hudson TJ, Tonin PN, Mes-Masson AM (2005) Discrimination between serous low malignant potential and invasive epithelial ovarian tumors using molecular profiling. Oncogene 24:4672–4687
Evangelou A, Jindal SK, Brown TJ, Letarte M (2000) Down-regulation of transforming growth factor beta receptors by androgen in ovarian cancer cells. Cancer Res 60:929–935
Hirte HW, Clark DA, Mazurka J, O’Connell G, Rusthoven J (1992) A rapid and simple method for the purification of tumor cells from ascitic fluid of ovarian carcinoma. Gynecol Oncol 44:223–226
Kruk PA, Maines-Bandiera SL, Auersperg N (1990) A simplified method to culture human ovarian surface epithelium. Lab Invest 63:132–136
Lawrenson K, Benjamin E, Turmaine M, Jacobs I, Gayther S, Dafou D (2009) In vitro three-dimensional modelling of human ovarian surface epithelial cells. Cell Prolif 42:385–393
Liu T, Cheng W, Lai D, Huang Y, Guo L (2010) Characterization of primary ovarian cancer cells in different culture systems. Oncol Rep 23:1277–1284
Lounis H, Provencher D, Godbout C, Fink D, Milot MJ, Mes-Masson AM (1994) Primary cultures of normal and tumoral human ovarian epithelium: a powerful tool for basic molecular studies. Exp Cell Res 215:303–309
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
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
Ouellet V, Zietarska M, Portelance L, Lafontaine J, Madore J, Puiffe ML, Arcand SL, Shen Z, Hebert J, Tonin PN, Provencher DM, Mes-Masson AM (2008) Characterization of three new serous epithelial ovarian cancer cell lines. BMC Cancer 8:152
Shepherd TG, Theriault BL, Campbell EJ, Nachtigal MW (2006) Primary culture of ovarian surface epithelial cells and ascites-derived ovarian cancer cells from patients. Nat Protoc 1:2643–2649
Onda T, Yoshikawa H (2011) Neoadjuvant chemotherapy for advanced ovarian cancer: overview of outcomes and unanswered questions. Expert Rev Anticancer Ther 11:1053–1067
Stewart JM, Shaw PA, Gedye C, Bernardini MQ, Neel BG, Ailles LE (2011) Phenotypic heterogeneity and instability of human ovarian tumor-initiating cells. Proc Natl Acad Sci U S A 108:6468–6473
Acknowledgments
Dr. Cécile Le Page for helpful suggestions. This work was supported by the banque de tissus et de données of the Fonds de Recherche Santé Québec (FRQS) and the Manitoba Ovarian Biobank Program (MOBP) through the Manitoba Tumor Bank and CancerCare Manitoba.
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Thériault, B.L., Portelance, L., Mes-Masson, AM., Nachtigal, M.W. (2013). Establishment of Primary Cultures from Ovarian Tumor Tissue and Ascites Fluid. In: Malek, A., Tchernitsa, O. (eds) Ovarian Cancer. Methods in Molecular Biology, vol 1049. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-547-7_24
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DOI: https://doi.org/10.1007/978-1-62703-547-7_24
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