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

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...

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Bibliographic Details
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
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Summary: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.
ISSN:1344-3941
1740-0929
DOI:10.1111/asj.13465