In vitro culture of primary human or mouse OSE often requires inclusion of insulin in the media to induce pro liferation. Although insulin and the related growth factor IGF I have been shown to alter epithelial polarity and directional cell growth, little is known about how these growth factors may affect directional growth of the OSE. Normal OSE grows on the outer surface of the ovary as a single layer of squamous to cuboidal epi thelium, however, at concentrations routinely used for culture of primary cells, insulin and IGF I induced for mation of hyperplastic OSE 4 6 cell layers thick likely due to a dramatic increase in the percentage of OSE undergoing proliferation. Importantly, the concentrations used in the present study and in typical cell culture media are higher than circulating levels or levels found in follicular fluid.

kinase inhibitor Physiological concentrations in the ovary range from 0. 5 10 ng mL in sulin and 100 500 ng mL IGF. Previously IGF1 at 100 ng mL was reported to increase OSE proliferation. The signaling pathway primarily responsible for this hyperplasia was the PI3K pathway, as inclusion of the PI3K inhibitor LY294002 restored growth of the OSE to a single cell layer. The PI3K pathway plays an important role in cell polarity through regula tion of the actin cytoskeleton. Activation of PI3K at the plasma membrane in turn leads to activation of Akt, which plays a critical role in chemotaxis and migration of many normal as well as cancerous cell types.

Ac tivation of this pathway may also repress expression of E cadherin, a component of the epithelial cell (-)-p-Bromotetramisole alkaline phosphatase inhibitor tight junc tion that functions to establish and maintain cell polarity that is often altered in ovarian cancer cells to permit increased metastasis. While no universally accepted precursor lesion exists for ovarian cancer originating in the OSE, menopausal ovaries and some mouse models of ovarian cancer exhibit hyperplasia of the OSE, forma tion of papillary structures, and inclusion cysts. Insulin and IGF I did not induce transformative changes in OSE as measured by growth in soft agar, however, it is possible that if levels of insulin and IGF accumulate enough locally in disease they might act on early stages of ovarian cancer to increase prolif eration and alter cell polarity to encourage hyperplasia. The OSE is able to secrete its own ECM, which may play a role in wound healing following ovulation.

In particular, OSE express collagen I and collagen IV in the basement membrane that delineates the OSE from the stroma. Since insulin and IGF I induced formation of hyperplastic OSE, the effects of insulin and IGF I on collagen IV expression and localization were analyzed to determine if the hyperplasia included changes in cell polarity. Organoids cultured in basal media exhib ited strong co