Figure 1

Study scheme. Scheme used for in vitro culture of mouse follicles for the assessment of follicle diameter, follicle survival, antral-like cavity formation, 17β-estradiol (E2) concentration in the culture media, ovulation, and germinal vesicle breakdown (GVBD) after human chorionic gonadotropin (hCG)treatment is shown. Early preantral follicles were mechanically isolated from mice ovaries as described in Methods. Collected follicles were individually cultured per well in a 24-well plate for 12 days. Follicles were cultured with various treatments. The follicles were inspected for morphology, follicle survival, and antral-like cavity formation every other day as described in Methods. Media were refreshed every other day, and E2 concentrations in the collected media were measured. After 12 days of culture, hCG (5 IU/mL) was added to the culture media to induce ovulation. After 16 h of hCG treatment, ovulation and GVBD were evaluated.