Dynamic of extracellular matrix remodeling and follicular morphology in cattle ovarian slices cultivated in medium supplemented with Croton argyrophyllus Kunth essential oil

CONTEXT: Ovarian tissue culture is commonly associated with increased follicular degeneration and extracellular matrix (ECM) changes largely owing to oxidative stress. AIMS: To investigate whether culture medium supplementation with Croton argyrophyllus Kunth essential oil (CAEO) attenuates the alterations in follicle morphology, integrity of ECM and stromal cells in cultured tissues. METHODS: Ovarian slices were cultivated for 6 days in α-MEM+ alone or with 0.01, 0.1, 1.0, 10.0, and 100.0 μg/mL CAEO. Subsequent to the culture period, the proportion of follicle survival and development were determined by histological analysis. The collagen and glycosaminoglycan areas, as well as the stromal cells numbers, were determined. KEY RESULTS: Ovarian tissue cultivated with 10.0 and 100.0 μg/mL CAEO showed higher percentage of normal follicles. Irrespective of the concentration, CAEO promoted an increase in collagen fiber density compared with tissues cultivated in α-MEM+ only, keeping the values similar to uncultured control. A reduction in collagen density was observed around primary follicles in comparison to primordial follicles in cultured tissues. The CAEO did not influence glycosaminoglycans, but the presence of 100.0 μg/mL CAEO increased the number of stromal cells compared with tissues cultured in control medium. CONCLUSIONS: The CAEO (10 and 100 μg/mL) increased follicle survival and improved collagen fiber organization in cultured ovarian slices. The stromal cell density was also increased in tissues cultured with 100.0 μg/mL CAEO. IMPLICATIONS: The CAEO is a promising supplement for improving in vitro ovarian tissue culture conditions.