Fig. 1

Study design describing the different conditions. Eight ovaries were cut into 2 parts and distributed among the four groups of transport media containing anti-apoptotic drugs (10 μM S1P or 10 μM Z-VAD-FMK) or vehicle (Control (CT); NaOH or DMSO). After the tissue was prepared, the ovarian cortex was cut into pieces with a biopsy punch to obtain similarly sized fragments (diameter of 2 mm). These pieces (24 punches per group), which were from different ovaries, were mixed and then randomly assigned to processing for the fresh histological analyses (6 punches) or to freezing (6 punches). After freezing and thawing, 6 punches were fixed (F-T), and the others were cultured for 2 or 6 days (D2 or D6). For histology, each entire punch was cut, and 12 sections were analyzed for follicular density and morphological assessment, whereas 3 sections were analyzed for proliferation