Figure 2.

Cell proliferation status in cultures of human testis tissue with areas containing full spermatogenesis, termed normal testis determined by KI-67 staining and BrdU incorporation after 0, 7 and 14 days of culture in the presence of 10% FBS. (A) Top panel shows IHC staining with KI-67. Bottom panel shows IHC staining with BrdU. BrdU staining detects incorporated BrdU during replication for 3 h before tissue fixation. Inserts show negative control staining (no antibody). Serial sections of tissue counterstained with Mayer haematoxylin are shown in each column. Tissue from six patients was investigated, with three experimental replicates per time point per patient. Scale bar corresponds to 50 μm. (B) Quantification of proliferating germ cells (spermatogonia and spermatocytes), determined as the percentage of KI-67 staining and BrdU incorporation. Values represent mean±s.e.m. * indicates significant difference (P<0.05).