Effect of centrifugal fractionation protocols on quality and recovery rate of equine sperm.
Abstract: Centrifugal fractionation of semen is commonly done to improve quality of human semen in assisted-reproduction laboratories, allowing sperm separation based on their isopycnic points. Sperm with morphologic abnormalities are often more buoyant, promoting their retention above defined density media, with structurally normal sperm passing through the media following centrifugation. Three experiments were conducted to evaluate the effects of density-medium type, centrifuge-tube size, sperm number, and density-medium volume (column height) on stallion sperm quality and recovery rate in sperm pellets following centrifugation. In all three experiments, equine semen was initially centrifuged to increase sperm concentration. In Experiment 1, semen was layered over continuous or discontinuous gradients. For Experiment 2, semen was layered over three column heights of continuous gradients in 15- or 50-ml conical-bottom tubes. For Experiment 3, increasing sperm numbers were layered over continuous gradient in 15- or 50-ml conical-bottom tubes. Following centrifugation, sperm pellets were evaluated for sperm morphologic quality, motility, DNA integrity, and recovery rate. Centrifugal fractionation improved (P < 0.05) sperm morphology, motility, and DNA integrity, as compared to controls. The continuous gradient increased (P < 0.05) sperm recovery rate relative to the discontinuous gradient, whereas sperm processed in 15-ml tubes yielded higher velocity and higher recovery rates (P 0.05) by column height of gradient. Increasing sperm number subjected to gradient centrifugation decreased (P < 0.05) sperm recovery rate when 15-ml tubes were used.
Copyright © 2012 Elsevier Inc. All rights reserved.
Publication Date: 2011-11-23 PubMed ID: 22115813DOI: 10.1016/j.theriogenology.2011.10.002Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study examines the impact of various centrifugal fractionation protocols on the quality and recovery rate of horse sperm. The researchers find that centrifugal fractionation enhances sperm morphology, motility, and DNA integrity compared to controls.
Introduction
The study was rooted in the common practice of centrifugal fractionation of semen which is usually used to enhance the quality of human sperm in assisted-reproduction labs. This process separates sperm based on their isopycnic points, thus isolating sperm with morphological abnormalities that are typically more buoyant. The normal sperm passes through the defined density media post-centrifugation.
Research Method
- The researchers conducted three experiments, during which they evaluated the effects of various factors such as the type of density medium, the size of the centrifuge tube, the number of sperm, and the volume of the density medium on the quality and recovery rate of stallion sperm post-centrifugation. The initial stage of each experiment involved centrifuging the equine semen to increase sperm concentration.
- In the first experiment, they layered semen over either continuous or discontinuous gradients.
- The second experiment involved layering semen over three column heights of continuous gradients in either 15- or 50-ml conical-bottom tubes.
- In the third experiment, they layered increasing numbers of sperm over a continuous gradient in 15- or 50-ml conical-bottom tubes.
Research Findings
- Upon centrifugation, the scientists evaluated the resultant sperm pellets for various quality aspects such as morphologic quality, motility, DNA integrity, and recovery rate. The findings revealed that centrifugal fractionation improved sperm morphology, motility, and DNA integrity compared to the controls.
- The use of a continuous gradient resulted in an elevated sperm recovery rate compared to discontinuous gradients.
- Sperm processed in tubes with a capacity of 15-ml yielded superior velocity and higher recovery rates compared to those processed in 50-ml tubes.
- The column height of the gradient did not impact the sperm recovery rate.
- When 15-ml tubes were used, increasing the number of sperm subjected to gradient centrifugation resulted in a decrease in the sperm recovery rate.
Cite This Article
APA
Edmond AJ, Brinsko SP, Love CC, Blanchard TL, Teague SR, Varner DD.
(2011).
Effect of centrifugal fractionation protocols on quality and recovery rate of equine sperm.
Theriogenology, 77(5), 959-966.
https://doi.org/10.1016/j.theriogenology.2011.10.002 Publication
Researcher Affiliations
- Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, Texas, USA.
MeSH Terms
- Animals
- Cell Separation / methods
- Cell Separation / veterinary
- Centrifugation, Density Gradient / methods
- Centrifugation, Density Gradient / veterinary
- DNA / analysis
- Horses
- Male
- Sperm Motility
- Spermatozoa / abnormalities
- Spermatozoa / chemistry
- Spermatozoa / cytology
Citations
This article has been cited 3 times.- Umair M, Henning H, Stout TAE, Claes A. A Modified Flotation Density Gradient Centrifugation Technique Improves the Semen Quality of Stallions with a High DNA Fragmentation Index. Animals (Basel) 2021 Jul 1;11(7).
- Gloria A, Carluccio A, Wegher L, Robbe D, Befacchia G, Contri A. Single and double layer centrifugation improve the quality of cryopreserved bovine sperm from poor quality ejaculates. J Anim Sci Biotechnol 2016;7:30.
- Inoue O, Kuji N, Ito H, Yamada M, Hamatani T, Oyadomari A, Kato S, Hanabusa H, Isaka K, Tanaka M. Clinical efficacy of a combination of Percoll continuous density gradient and swim-up techniques for semen processing in HIV-1 serodiscordant couples. Asian J Androl 2017 Mar-Apr;19(2):208-213.
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