Effect of ovarian storage time at 4 degrees C on cumulus cell apoptosis in porcine antral follicles

• Published in: Animal science journal 2020-01, Vol.91 (1), p.e13465-n/a
• Main Authors: Chi, Yafei, Peng, Boya, Lu, Jing
• Format: Article
• Language: English
• Subjects: Animals; Annexin V; Annexin V staining; Apoptosis; Bioassays; Cell Separation; Cells, Cultured; Cold storage; Cold Temperature - adverse effects; Comet assay; cumulus cell; Cumulus Cells - pathology; Cumulus Cells - physiology; Damage detection; Deoxyribonucleic acid; DNA; DNA nucleotidylexotransferase; Female; Follicles; Granulosa cells; In Situ Nick-End Labeling - methods; Organ Preservation - adverse effects; Organ Preservation - methods; Ovarian Follicle - cytology; Ovaries; ovary; Ovary - cytology; Physiology; Saline solutions; Staining; Swine; Time dependence; Time Factors
• ISSN: 1344-3941; 1740-0929
• DOI: 10.1111/asj.13465

The present study was conducted to investigate the effect of cold storage time on apoptosis of cumulus cells (CCs) from porcine ovaries, and to compare the sensitivity of four apoptosis‐detection methods. Porcine ovaries were stored in physiological saline solution at 4°C for 0, 7, 24 and 48 hr, and then cumulus cells or granulosa cells (GCs) in antral follicles were retrieved to detect cell apoptosis. Cumulus cells isolated from stored ovaries for 24 hr presented obvious apoptosis using terminal deoxynucleotidyl transferase (TdT)‐mediated d‐UTP nick end‐labeling (TUNEL) assay. A typical DNA ladder pattern of apoptosis was observed in GCs 24 hr post storage treatment. The mean Olive Tail Moment of CCs was significantly increased after 24 hr using comet assay; however, the mean tail migration and mean tail DNA increased gradually after 7 hr of storage. In addition, annexin V/PI staining assay showed an obvious increase in apoptotic CCs (Annexin V positive, PI negative) 7 hr after treatment, and the apoptotic rate reached to a peak at 24 hr followed by a decline after 48 hr of storage to the level at 7 hr. In conclusion, cold storage of porcine ovary in physiological saline solution induced a time‐dependent increase in apoptosis of cumulus cells, and annexin V/PI staining combined with comet assay provided a sensitive and reliable method to detect early damages in cumulus cells induced by cold storage of ovary.