Co-Cultivation of Human Granulosa Cells with Ovarian Cancer Cells Leads to a Significant Increase in Progesterone Production

Abstract

Abstract In humans, granulosa cells (GCs) are part of the follicle and nourish the growing oocyte. GCs produce oestrogen and, after ovulation, progesterone. They are embedded in a multicellular tissue structure of the ovary, which consists of a variety of different cell types that are essential for the physiological function of the ovary. However, the extent to which individual ovarian cell types contribute to overall functionality has not yet been fully elucidated. In this study, we aim to investigate the effects of co-culturing human granulosa cells with ovarian cancer cells on their progesterone and oestrogen production in an in vitro model. After seeding, the cells were stimulated with 200µM forskolin in DMEM for 72 hours and the medium of the different cell culture experiments was collected. Subsequently, progesterone and oestradiol concentrations were determined using an Elisa assay. Morphologically, it was striking that the cells self-organise and form spatially separated areas. Compared to culturing granulosa cells alone, co-culturing human granulosa cells together with the ovarian cancer cell line OvCar-3 resulted in a significant increase in progesterone production (20.3ng/ml versus 50.2ng/ml; p < 0.01). Using a simple in vitro model, we highlight the importance of cellular crosstalk between different ovarian cells in a complex cellular network and that it strongly influences granulosa cell hormone production. This could have potential implications for the procedure of transplanting endocrine tissues after cryopreservation, as it highlights the importance of survival of all cells for the functionality of the transplanted tissue.