Abstract: A total of 88 ovaries in the middle follicular stage were used to detect apoptosis using the modified staining procedure with terminal deoxynucleotide transferase-mediated dUTP nick end-labeling assay (TUNEL), Hematoxylin and Eosin. The analysis observed proportions of cells for apoptosis with several options of culture systems, presenting a control (77.2±7.0), culture 1 (3.8±2.0), culture 2 (1.3±2.0), culture 3 (72.7±6.0), culture 4 (6.7±5.0), culture 5 (80.1±6.0) and culture 6 (80.2±6.0). The apoptotic cells in cultures 1, 2 and 4 were significantly different with other cultures including the control whereas cultures 3, 5 and 6 showed significantly similar apoptotic patterns with the control. The present analysis observed clear differences for relative numbers of internal ovary cells in apoptosis and necrosis. The new circulation method provided from this study with culture media, levels of gas supply (O2 and CO2) and incubation temperature for in vitro culture systems can prolong the lifetime of cells.