Apoptosis quantification by TUNEL analysis.

<p>(A) cell nuclei stained with DAPI (blue) to perform the TUNEL assay. The sample from the G3 group. The TUNEL + signal was found in the stoma cells (red) from G3; (B) note the primordial follicle in the G2 sample. The TUNEL + signal was found in the stroma, but note that it was absent in the oocyte and GC from the follicles of the G2 group; (C) the G6 sample. Primordial follicles surrounded by flattened shaped GC and stroma nuclei stained with DAPI (blue). TUNEL-positive cells stained in red in the stroma. Follicles were negative for DNA damage in the G6 sample; (D) the G5 sample that contained two primordial follicles, cell nuclei stained with DAPI. No TUNEL + signal was detected in oocyte and GC, but was positive in the ovarian stroma; (E) cell death index. Quantification of the follicles with a positive TUNEL signal in oocytes or GC. No differences were detected between activated and non activated samples; (F) TUNEL-positive stromal area quantification. No differences were detected for follicles when the TUNEL+ area was compared between the activated and non activated groups. This finding suggests that the cell damage found in the stroma from all the groups was due to the culture process.