Organotypic culture of human ovarian surface epithelial cells: A potential model for ovarian carcinogenesis

Abstract

The objective of this work was to establish an in vitro multidimensional culture system for human ovarian surface epithelial (HOSE) cells as a model for ovarian carcinogenesis. The epithelial origin of cell outgrowth from cells obtained from the ovarian surface was confirmed by keratin staining. Two cultures from two different patients were established, HOSE-A and HOSE-B. Cultures were infected with a retrovirus expressing human papillomavirus genes E6 and E7 to extend their life span. HOSE cells were seeded onto collagen gels containing NIH3T3-J2 fibroblasts as feeder cells and grown to confluence submerged in growth medium. The collagen bed was then raised to the air-medium interface for 7 d (organotypic culture). Microscopically, fixed cultures revealed a single layer of flat cells growing on the collagen surface, reminiscent of HOSE cells in vivo. Infected HOSE-A and HOSE-B cells exhibited aberrant growth because they stratified. In addition, established ovarian cancer lines grown in this fashion stratified and showed malignant phenotypes. Thus, cells grown in organotypic culture resemble their in vivo counterparts, providing a basis for establishing a system to study growth, proliferation, differential gene expression, and perhaps malignant transformation of HOSE cells.

Reversion of transformed phenotype in ovarian cancer cells by intracellular expression of anti folate receptor antibodies

“…We placed the collagen bed at the air-medium interface since it more closely resembles normal ovary structure and discriminates tridimensional growth of normal OSE cells from that of immortalized OSE or malignant ovarian carcinoma lines. 31 FR downmodulated cells tended toward ordered growth in a single or few layers in tridimensional cultures, suggesting that the increase in a6b4 expression might lead to a restoration of the negative signaling of this integrin. 32 The inability of clone DM99 to form multilayers in organotypic culture, despite the high folic acid content in the medium, suggests a role for FR in cell de-differentiation rather than in cell proliferation.…”

“…30,31 While monolayer cultures are suited for evaluation of growth rates, they do not recapitulate interactions with similar and different cell types of cells and with extracellular matrix. We placed the collagen bed at the air-medium interface since it more closely resembles normal ovary structure and discriminates tridimensional growth of normal OSE cells from that of immortalized OSE or malignant ovarian carcinoma lines.…”

Crucial Roles of Sp1 and Epigenetic Modifications in the Regulation of the CLDN4 Promoter in Ovarian Cancer Cells

“…Ovarian cancer cell line A2780 was cultivated in RPMI1640 (Invitrogen) supplemented with 10% FBS, 4 g/ml human recombinant insulin (Invitrogen), and antibiotics. HOSE-B, an ovarian surface epithelial cell line immortalized with E6 and E7 (18), was cultivated in RPMI1640 supplemented with 10% FBS and antibiotics. ML-3, an SV40-immortalized ovarian cystadenoma cell line (19), was cultivated in McCoy's supplemented with 10% FBS and antibiotics.…”

Phosphorylation of Claudin-3 at Threonine 192 by cAMP-dependent Protein Kinase Regulates Tight Junction Barrier Function in Ovarian Cancer Cells

“…HOSE-B, an ovarian surface epithelial cell line immortalized with E6 and E7 (26), was maintained in RPMI 1640 medium supplemented with 10% fetal calf serum and 300 g/ml Geneticin (Invitrogen). HEK293 cells were maintained in McCoy's growth me-dium with the supplements described above.…”