Successful vitrification of manually punctured equine embryos.
Authors: Wilsher Sandra, Rigali Florencia, Kovacsy Sofia, Allen Wr Twink
Journal: Equine veterinary journal
Summary
# Editorial Summary: Manual Puncture Vitrification of Equine Embryos Blastocoele collapse has traditionally required expensive micromanipulator equipment to prepare equine expanded blastocysts for vitrification, limiting access to embryo freezing technology across the industry. Researchers vitrified 79 Day 7–8 embryos using commercial human vitrification kits (containing DMSO and ethylene glycol cryoprotectants with sucrose for warming), comparing outcomes between micromanipulator-assisted and simple manual microneedle puncture techniques across embryos of varying sizes and equilibration durations. Manually punctured embryos measuring ≤560 µm exposed to the equilibration solution for 6 minutes yielded an impressive 82% pregnancy rate (14/17), matching the 80% success rate of unpunctured embryos ≤300 µm, whilst larger embryos and extended equilibration periods resulted in significantly reduced viability. This straightforward approach—requiring only basic equipment and commercial kits designed for human use—offers equine practitioners a practical, cost-effective alternative to micromanipulator-dependent methods, potentially democratising access to embryo cryopreservation; however, strict adherence to embryo size parameters and timing protocols is essential to replicate these results clinically.
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Practical Takeaways
- •Manual puncture of equine embryos ≤560 µm using a microneedle is a cost-effective alternative to micromanipulator-assisted collapse, eliminating the need for expensive equipment
- •Optimal protocol involves 6 minutes exposure to equilibration solution; longer exposure times (8 minutes) reduce pregnancy success rates
- •This technique makes embryo vitrification more accessible to equine reproduction programs by reducing equipment costs while maintaining competitive pregnancy rates (80-82%)
Key Findings
- •Manually punctured equine embryos ≤560 µm exposed to equilibration solution for 6 minutes achieved 82% pregnancy rate (14/17)
- •Unpunctured embryos ≤300 µm achieved 80% pregnancy rate (8/10)
- •Larger unpunctured embryos (>300-≤560 µm), punctured large embryos (>560 µm), and embryos exposed to equilibration solution for 8 minutes showed significantly reduced pregnancy rates
- •Commercial human vitrification and warming kits can successfully be used for equine embryo cryopreservation with manual puncture technique