Cushioned versus noncushioned centrifugation: sperm recovery rate and integrity.
Abstract: It was hypothesized that optimal sperm recovery rate (RR) without damage to the sperm would be obtained after centrifugation without a cushion solution. Semen collected three times from six light breed stallions was extended to 25 × 10(6) sperm/mL and centrifuged at CON (noncentrifuged), 900NC (no-cushion), 900C (cushion), 1800NC, and 1800C × g for 10 minutes. Sperm concentration, motility (TM and PM), and intact plasma membranes (PLM) and acrosomes (ACR) pre- and postcentrifugation (D0) and after 24 hours (D1) of cooling were evaluated. The RR in the CON (100 ± 0.0), 900NC (93.7 ± 2.9), and 1800NC (96.7 ± 2.6) groups was significantly higher than the 900C (68.7 ± 4.6) and 1800C (79.6 ± 3.5) groups. The D0 TM and PM were not different between the CON, 900NC, 900C, and 1800C, but were lower for the 1800NC group. The D1 TM and PM of the 900NC (75.2 ± 3.8 and 71.1 ± 4.1) and 900C (76.2 ± 3.7 and 72.4 ± 4.0) groups were significantly higher than the 1800NC (71.7 ± 4.1 and 67.3 ± 4.4) and 1800C (71.6 ± 4.1 and 67.2 ± 4.4) groups, and the CON (66.2 ± 4.5 and 60.0 ± 4.8) group was significantly lower than the other groups. The D1 PLM of the CON, 900NC, 900C, 1800NC, and 1800C groups were not different. The ACR on D1 was significantly lower for the CON (93.0 ± 2.4) group compared with all other groups. Optimal RR preserving sperm integrity was obtained in the 900NC group.
Copyright © 2013 Elsevier Inc. All rights reserved.
Publication Date: 2013-07-09 PubMed ID: 23849256DOI: 10.1016/j.theriogenology.2013.06.009Google Scholar: Lookup
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- Comparative Study
- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article investigates the best method for sperm sample preparation, comparing procedures involving cushioned and non-cushioned centrifugation. The study found that non-cushioned centrifugation yielded a higher sperm recovery rate and better preservation of sperm integrity.
Research Techniques and Aims
- The researchers wanted to determine which centrifugation method – cushioned or non-cushioned – would yield the highest sperm recovery rate (RR) and preserve the integrity of the sperm cells.
- To this end, three rounds of semen samples were collected from six stallions of a light breed. These samples were diluted to a concentration of 25 million sperm cells per milliliter.
- The samples were then subjected to different centrifugation processes, including control (no centrifugation), 900 times gravity with no cushion (900NC), 900 times gravity with cushion (900C), 1800 times gravity with no cushion (1800NC), and 1800 times gravity with cushion (1800C).
Processes and Measurements
- The centrifugation took place over a span of 10 minutes. Before and after centrifugation (Day 0), and after a cooling period of 24 hours (Day 1), the researchers assessed sperm concentration, movement (both total motility (TM) and progressive motility (PM)), and intactness of sperm cell membranes (PLM) and cap structures (ACR) used in fertilization.
Results and Findings
- The findings showed that the sperm recovery rates in the control group (no centrifugation), the 900NC, and the 1800NC process were significantly higher than in the cushioned centrifugation processes (both 900C and 1800C).
- On Day 0, the total and progressive motility were not significantly different among the control, 900NC, 900C, and 1800C groups. However, these parameters were found to be lower in the 1800NC group.
- By Day 1, the total and progressive motility of the 900NC and 900C groups were significantly better than those of both the 1800NC and 1800C groups. The control group had the lowest values when compared to all other groups.
- However, the intactness of the sperm plasma membranes on Day 1 was not significantly different among all the groups.
- The integrity of acrosomes on Day 1 was significantly worse in the control group compared with the other centrifugation methods.
- Thus, the best sperm recovery rate and preservation of sperm integrity were found in the 900NC group, i.e., samples that were centrifuged at 900 times gravity without a cushion solution.
Cite This Article
APA
Len JA, Beehan DP, Lyle SK, Eilts BE.
(2013).
Cushioned versus noncushioned centrifugation: sperm recovery rate and integrity.
Theriogenology, 80(6), 648-653.
https://doi.org/10.1016/j.theriogenology.2013.06.009 Publication
Researcher Affiliations
- Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana, USA. jose.lenyin@adelaide.edu.au
MeSH Terms
- Acrosome / physiology
- Animals
- Cell Membrane / physiology
- Cell Separation / methods
- Cell Separation / statistics & numerical data
- Cell Separation / veterinary
- Centrifugation / methods
- Centrifugation / veterinary
- Horses
- Male
- Semen / cytology
- Semen Analysis / methods
- Semen Analysis / statistics & numerical data
- Semen Analysis / veterinary
- Sperm Count / statistics & numerical data
Citations
This article has been cited 2 times.- Marzano G, Moscatelli N, Di Giacomo M, Martino NA, Lacalandra GM, Dell'Aquila ME, Maruccio G, Primiceri E, Chiriacò MS, Zara V, Ferramosca A. Centrifugation Force and Time Alter CASA Parameters and Oxidative Status of Cryopreserved Stallion Sperm. Biology (Basel) 2020 Jan 27;9(2).
- 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.
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