Evaluation of viability and apoptosis in horse embryos stored under different conditions at 5 °C
| Autor: | Peter Daels, M. Moussa, J. L. Tremoleda, G. Duchamp, Ben Colenbrander, J.-F. Bruyas, M.M. Bevers |
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| Rok vydání: | 2004 |
| Předmět: |
Programmed cell death
Indoles Time Factors Necrosis Apoptosis DNA Fragmentation Biology chemistry.chemical_compound Ovulation Induction Food Animals In Situ Nick-End Labeling medicine Animals Horses DAPI Small Animals Insemination Artificial Fluorescent Dyes TUNEL assay Equine Embryo Embryo Mammalian Molecular biology eye diseases Staining Cold Temperature chemistry Tissue and Organ Harvesting DNA fragmentation Female Animal Science and Zoology Tissue Preservation sense organs medicine.symptom |
| Zdroj: | Theriogenology. 61:921-932 |
| ISSN: | 0093-691X |
| Popis: | The aim of this study was to evaluate the viability (percentage of dead cells) and the incidence of DNA fragmentation of horse embryos after storage in three different media at 5 degrees C for 6 and 24 h. Forty embryos were stored in Emcare Holding Solution for 6 and 24 h, in Hams'F10 or Vigro Holding Plus for 24 h at 5 degrees C (n = 9-10 per group) and 10 embryos were evaluated immediately after collection. First, embryos were stained, immediately after collection or following storage, to detect dead cells (DAPI) and, subsequently, DAPI-stained embryos were fixed and stained to detect DNA fragmentation (TUNEL). Finally, all the fixed embryos were re-stained with DAPI to determine the total number of cells. The percentage of cells stained with both TUNEL and DAPI or TUNEL-only or DAPI-only were determined. The percent of dead cells (DAPI-labelled) per embryo increased with duration of storage, but no differences were detected between the storage media. The percentage of early apoptotic cells (TUNEL+/DAPI-) in fresh and stored embryo for 6 h or 24 h did not differ significantly (P0.05). There was a significant correlation between the percentage of cells labelled by TUNEL and DAPI (R = 0.87) (P0.001). These results suggest that cooled storage increases cell death but this does not appear to occur by induction of apoptosis and that DAPI staining proves to be a quick and reliable method for assessing embryo viability. |
| Databáze: | OpenAIRE |
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