Serial cultivation of normal human ovarian surface epithelium in tissue culture : a potential model for ovarian carcinogenesis [article]

Craig Henry Siemens
2010
Most human ovarian cancers are thought to arise in the surface epithelium of the ovary. However, few models for the study of this tissue exist and no methods for its propagation in culture are available. This report addresses two problems encountered previously in the culture of normal human ovarian surface epithelium (OSE): (i) limited growth; and (ii) difficulties in cell identification because of the pleomorphism of cultured human OSE cells. The poor growth potential exhibited by human OSE
more » ... culture was greatly improved by the development of a method that permits, for the first time, the serial cultivation and clonal growth of human OSE in culture. To achieve this, OSE cells were cultured from premenopausal ovarian tissue obtained at surgery. Growth of OSE cells was compared in several culture media: Medium 199 (199), MCDB 202 (202), Waymouth's 752/1 (WM), and 199/202 (1:1), with 5, 15, or 25% fetal bovine serum (FBS) and with/without epidermal growth factor (EGF, 20 ng/ml) and hydrocortisone (HC, 0.4 μg/ml). Proliferation was greater in 199/202 (1:1) with 15$ FBS, EGF, and HC, than in the same medium with 5% FBS or Waymouth's medium with any supplement. At clonal densities (100 cells/60 mm culture dish) with EGF/HC, seeding efficiencies in most media ranged from 30 to 50%. Cloning efficiencies differed significantly between media and ranged from 1.0% to 13%. In the medium optimal for maximal proliferation, 15% FBS/199/202 (1:1)/EGF/HC, OSE cells were subcultured up to 10 times and underwent 20 to 25 population doublings. The population doubling time during the log phase of growth was approximately 48 hours. Seeding efficiencies of up to 53% and cloning efficiencies of up to 13% were obtained. The radii of outgrowths were significantly greater than in the other media. Early passage OSE cells reversibly modulated from an epithelial morphology, in medium lacking EGF/HC, to an elongate morphology in medium containing EGF/HC. The cytokeratin filaments of OSE modulated from a strongly-stained, filamentous patte [...]
doi:10.14288/1.0096807 fatcat:oowby3jnozcc5a2v2ual7fut3e