ORGANOTYPIC CULTURE OF HUMAN OVARIAN SURFACE EPITHELIAL-CELLS - A POTENTIAL MODEL FOR OVARIAN CARCINOGENESIS

The objective of this work was to establish an in vitro multidimension al culture system for human ovarian surface epithelial (HOSE) cells as a model for ovarian carcinogenesis. The epithelial origin of cell out growth from cells obtained from the ovarian surface was confirmed by k eratin staining. Two cultures from two different patients were establi shed, HOSE-A and HOSE-B. Cultures were infected with a retrovirus expr essing human papillomavirus genes E6 and E7 to extend their life span. HOSE cells were seeded onto collagen gels containing NIH3T3-J2 fibrob lasts as feeder cells and grown to confluence submerged in growth medi um. The collagen bed was then raised to the air-medium interface for 7 d (organotypic culture). Microscopically, fu;ed cultures revealed a s ingle layer of flat cells growing on the collagen surface, reminiscent of HOSE cells in vivo. Infected HOSE-A and HOSE-B cells exhibited abe rrant growth because they stratified. In addition, established ovarian cancer lines grown in this fashion stratified and showed malignant ph enotypes. Thus, cells grown in organotypic culture resemble their in v ivo counterparts, providing a basis for establishing a system to study growth, proliferation, differential gene expression, and perhaps mali gnant transformation of HOSE cells.