Effects of tumour necrosis factor‐alpha and interleukin‐1 beta on in vitro development of bovine secondary follicles

The objective of this study was to determine the effects of TNF‐α and IL‐1β on development and survival of bovine secondary follicle culture in vitro for 18 days. Secondary follicles (~0.2 mm) were isolated from ovarian cortex and individually cultured at 38.5°C, with 5% CO2 in air, for 18 days, in TCM‐199+ alone (cultured control) or supplemented with 10 ng/ml IL‐1β, 10 ng/ml TNF‐α or both TNF‐α and IL‐1β. The effects of these treatments on growth, follicular survival, antrum formation, viability, ultrastructure and mRNA levels for GDF‐9, c‐MOS, H1foo and Cyclin B1 were evaluated. The results showed that addition of TNF‐α to culture medium increased follicular diameter and rate of antrum formation, whereas that of IL‐1β and a mixture of IL‐1β and TNF‐α did not do so. Ultrastructural analysis showed that, among the tested cytokine treatments, follicles cultured in the presence of TNF‐α had the best‐preserved oocytes and granulosa cells. The presence of TNF‐α, IL‐1β or both did not influence the expression of mRNAs analysed. In conclusion, in contrast to IL‐1β, TNF‐α promotes growth of and antrum formation in in vitro cultured bovine secondary follicles, while their ultrastructure and viability were maintained.