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 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.
Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
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L
Gregoire
A
A
Munkarah
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R
Rabah
R T
RT
Morris
W D
WD
Lancaster
D016475:3T3 Cells; D000818:Animals; D018929:Cell Culture Techniques; D002455:Cell Division; D002471:Cell Transformation, Neoplastic; D004847:Epithelial Cells; D005260:Female; D005814:Genes, Viral; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008954:Models, Biological; D009856:Oncogene Proteins, Viral / Q000235:genetics; D010051:Ovarian Neoplasms / Q000473:pathology*; D010053:Ovary / Q000166:cytology*; D027383:Papillomaviridae / Q000235:genetics; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; D012190:Retroviridae / Q000235:genetics
C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16