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Sperm Collection of Differential Quality Using Density Gradient Centrifugation
Published on: November 29, 2018
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Sperm sexing with density gradient centrifugation in dogs
Gabriele Barros Mothé1, Caroline Scott1, Carmen Cecília Sicherle1
1Department of Animal Reproduction and Veterinary Radiology, College of Veterinary Medicine and Animal Science, São Paulo State University, Botucatu, São Paulo, Brazil.
Animal Reproduction Science
|November 21, 2018
Summary
Sexing dog sperm using density gradients was evaluated for commercial application. None of the tested methods effectively separated X and Y chromosome-containing sperm, indicating current techniques are not commercially viable.
Area of Science:
- Animal Reproduction
- Sperm Sexing Technologies
- Canine Reproduction
Background:
- Polytocous species like dogs present market opportunities for same-sex offspring.
- Limited research exists on sperm sexing effectiveness in canines.
- Improving canine breeding efficiency through sexed semen is a significant goal.
Purpose of the Study:
- To assess the efficacy of three discontinuous density gradients for separating X and Y chromosome-bearing dog sperm.
- To evaluate the impact of these separation methods on overall sperm quality.
Main Methods:
- Sperm sexing was attempted using Percoll®, Percoll®/Nycodenz®, and Percoll®/Ficoll gradients.
- Sperm quality parameters including motility, concentration, morphology, and membrane integrity were assessed.
- Quantitative real-time PCR (qPCR) was used to determine the percentage of X and Y chromosome-carrying sperm pre- and post-separation.
Main Results:
- The Ficoll gradient yielded the best post-centrifugation sperm quality.
- No significant enrichment of X or Y chromosome-containing sperm was achieved with any gradient method.
- Percentages of X and Y sperm remained largely unchanged post-gradient separation across all methods.
Conclusions:
- Density gradient centrifugation techniques are not effective for sexing dog sperm for commercial purposes.
- Further research is needed to develop viable sperm sexing methods for canines.
- Current sexing technologies do not meet the requirements for commercial application in dogs.
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