Optimizing equine sperm quality: an alternative to single layer centrifugation for sperm isolation

Area of Science:

• Reproductive Biology
• Animal Science
• Biotechnology

Background:

• In vitro semen purification aims to mimic in vivo selection for optimal oocyte fertilization.
• Single-layer centrifugation (e.g., EquiPure) is the established method for isolating functional spermatozoa for artificial insemination.
• Existing methods often involve complex procedures, necessitating simpler and more effective alternatives.

Purpose of the Study:

• To introduce and evaluate a novel AI port system for spermatozoa isolation.
• To compare the efficacy of the AI port system against the commercial EquiPure method.
• To assess the impact of the AI port system on key sperm quality parameters.

Main Methods:

• The AI port system utilizes a disposable cartridge with two chambers separated by a 5 µm polyethylene terephthalate (PETE) membrane.
• Highly motile spermatozoa migrate across the membrane over 20 minutes, leaving behind seminal plasma and other cells.
• Spermatozoa isolated via the AI port system were compared to those isolated using EquiPure.

Main Results:

• The AI port system demonstrated comparable or superior performance across all tested sperm parameters: yield, vitality, motility, morphology, DNA fragmentation, and mitochondrial superoxide generation.
• Notably, AI port-isolated spermatozoa showed significantly lower DNA fragmentation.
• The system effectively removed seminal plasma and leukocytes.

Conclusions:

• The AI port system is a potentially cost-effective and user-friendly alternative for isolating high-quality spermatozoa.
• This novel method may enhance artificial insemination success rates in equine breeding programs.
• The AI port system shows promise for efficient and effective sperm purification.