Trimethylation profile of histones H3 lysine 4 and 9 in late preantral and early antral caprine follicles grown in vivo versus in vitro in the presence of anethole.

This study assessed the histones methylation profile (H3K4me3 and H3K9me3) in late preantral (PA) and early antral (EA) caprine follicles grown in vivo and in vitro, and the anethole effect during in vitro culture of PA follicles. Uncultured in vivo-grown follicles (PA, n = 64; EA, n = 73) were used as controls to assess the methylation profile and genes' expression related to apoptosis cascade (BAX, proapoptotic; BCL2, antiapoptotic), steroidogenesis (CYP17, CYP19A1), and demethylation (KDM1AX1, KDM1AX2, KDM3A). The isolated PA follicles (n = 174) were cultured in vitro for 6 days in α-MEM+ in either absence (control) or presence of anethole. After culture, EA follicles were evaluated for methylation, mRNA abundance, and morphometry. Follicle diameter increased after culture, regardless of treatment. The methylation profile and the mRNA abundance were similar between in vivo-grown PA and EA follicles. Anethole treatment led to higher H3K4me3 fluorescence intensity in EA follicles. The mRNA abundances of BAX, CYP17, and CYP19A1 were higher, and BCL2 and KDM3A were lower in in vitro-grown EA follicles than in vivo-grown follicles. In conclusion, in vitro follicle culture affected H3K4me3 fluorescence intensity, mRNA abundance of apoptotic genes, and steroidogenic and demethylase enzymes compared with in vivo-grown follicles.

Improving survival and growth of caprine preantral follicles cultured in medium commonly used for MSC: Role of oxidative stress regulation and epigenetic changes.

This study evaluated the efficiency of in vitro culture of preantral follicles (PAF) in a commonly used medium for mesenchymal stem cell (MSC) culture. Parameters assessed included follicle survival, growth, stromal cell density, levels of reduced thiols and reactive oxygen species, epigenetic changes, cell apoptosis, and mRNA abundance. Caprine ovarian tissues were cultured for 1 or 7 days in either PAF or MSC-common media, with uncultured tissues serving as controls. The MSC medium exhibited increased follicular survival and growth and remodeled stromal density potentially through the regulation of oxidative stress and epigenetic changes compared to the PAF medium. In conclusion, our results highlight the importance of the MSC medium in enhancing follicular survival and growth, changing the stromal cell density, as well as in regulating the medium oxidative stress and epigenetic changes during the in vitro culture of caprine PAF.