Effect of embryo age on the viability of equine embryos after cooled storage using two transport systems

The aim of this study was to compare the viability of 7- and 8-day-old equine embryos cooled and stored for 6 or 24 hours in two different transport systems. Embryos (n = 97) were recovered on day 7 or 8 and assigned to 10 groups (n = 10/group). Embryos within the same age group (D7 or D8) were evaluated immediately after collection (Group-0h) or after storage in an Equitainer at 5°C for 24 hours in 5 ml Emcare Holding Solution (EHS) (Group-E-24h) or 5 ml Ham's F10 (Group-H-24h) or in a refrigerator at 5°C in 500 ml Emcare Flushing Solution (EFS) for 6 hours (Group-B-6h) or 24 hours (Group-B-24h). After collection or storage, embryos were incubated in 1 μg/ml DAPI to determine the percentage of dead cells per embryo (DAPI positive, fluorescent cells). Subsequently, embryos were fixed in 4% paraformaldehyde and re-stained with DAPI to determine the total number of cells. The percentage of dead cells in group-0h and B-6h was similar and significantly lower than for embryos stored for 24 hours in groups B-24h, E-24h, and H-24h. The percentage of dead cells was similar for embryos stored in an Equitainer (groups E-24h and H-24h) and was significantly higher for embryos stored 24 hours in EFS (Group B-24h). Within each storage system (0h, B-6h, B-24h, E-24h, and H-24h) no significant difference in the percentage of dead cells was observed between 7- and 8-day-old embryos. Storage in 500 ml EFS at 5°C for 6 hours resulted in embryos of better quality than after the traditional 24-hour storage in an Equitainer, suggesting that this simplified system offers a good alternative for short-term storage and transport.