Background: This study aimed to select high-quality spermatozoa by fresh domestic feline semen using sperm separation by magnetic activation (MASS), compared to density gradient centrifugation (DGC), and to evaluate cell quality after selection. Methods: Semen was collected from ten domestic felines by pharmacological sampling using dexmedetomidine and ketamine followed by urethral catheterization. The following parameters were analyzed: motility (computer assisted semen analysis), concentration (Neubauer chamber), semen morphology (humid chamber), and supravital test (eosin/nigrosine staining). In DGC, 20 × 106 spermatozoa were used in a gradient of Percoll at 90% and 45%, centrifuged at 900 g for 5 min, and the pellet was diluted in HEPES buffer. In MASS, 10 × 106 spermatozoa were diluted in HEPES buffer, centrifuged at 300 g for 10 min. The pellet was then resuspended in HEPES buffer with nanoparticles bound to annexin V, incubated for 15 min, and then filtered in the magnetic separation column. Results: The total abnormalities in the fresh semen were 47.9 ± 4.47%, with DGC and MASS being effective in reducing sperm abnormalities by 15.4 ± 0.95% and 29.80 ± 4.90%, respectively. Conclusions: This nanotechnological method is efficient in producing high-quality semen samples for assisted reproduction.
목차
ABSTRACT INTRODUCTION MATERIALS AND METHODS Ethical aspect The animals Semen collection and analysis Density gradient centrifugation Sperm separation of magnetic activation Statistical analysis RESULTS DISCUSSION CONCLUSION REFERENCES
Teresinha Inês Assumpção [ Laboratory of Animal Reproduction, Faculty of Veterinary Medicine and Animal Science, Federal University of Uberlândia, Uberlândia 38410337, Brazil ]
Corresponding Author
Maria Isabel Mello Martins [ Department of Veterinary Clinics, State University of Londrina, Londrina 86057970, Brazil ]
Caroline Silva Vieira [ Reproduction Physiology Researcher, Reneco International Wildlife Consultants, Abu Dhabi 61741, UAE ]
Mauricio Foschini [ Physics Institute, Federal University of Uberlândia, Uberlândia 38408100, Brazil ]