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Updated: Aug 8, 2026
Enrichment of Pachytene Spermatocytes and Spermatids from Mouse Testes Using Standard Laboratory Equipment
Published on: September 17, 2019
Improvements and short-term viability of mouse epididymal spermatozoa recovered through the Spermprep filtration
P M Zavos1, J R Correa, P N Zarmakoupis
1Department of Animal Sciences, University of Kentucky, Lexington, KY 40546, USA.
Abstract:
This study was designed to determine the effects of Sephadex filtration (SpermprepI method) on the separation of motile, morphologically normal, mouse epididymal spermatozoa and to study the viability of the recovered spermatozoa over a 3-h incubation period. Spermatozoa were harvested from the caudae epididymie (5 animals per run or replication; n=10) following bilateral testicular excision, after which they were incubated in 2-ml of Test-Yolk buffer (TYB) at 37 degrees C for 15-min. The specimens were then split into 2 1-ml aliquots, with Aliquot 1 as the control and Aliquot 2 as the filtered sample. The SpermprepI column was employed according to the manufacturer's specifications using TYB. During filtration (10-min), 2 different fractions were obtained: first 5-min (Sample 1) and second 5-min (Sample 2). The 2 fractions were evaluated and incubated at 37 degrees C and assessed for percentage of motility and grade of motility (0 to 4) every 30-min for 3-h. Filtration resulted in a significant improvement in the percentage and grade of motility (91.5% and 3.0 vs 76.5% and 2.5, respectively). The results indicate that filtration with the SpermprepI method improved the percentage and grade of motility (P<0.05) but not the percentage of normal morphology of the spermatozoa. In addition, the SpermprepI method enabled the recovery of 45% (8.3x10(6) spermatozoa recovered) of the total number of spermatozoa processed in the control aliquot (18.4x10(6) spermatozoa), which is consistent with previous observations. Most importantly, filtered spermatozoa incubated for 3-h showed a greater percentage and grade of motility than the control spermatozoa (63% and 1.66 vs 39% and 0.82, respectively. The SpermprepI filtration method selected a higher proportion of quality spermatozoa, which also displayed significant long-term motility (longevity) during in vitro incubation.
Insights
Sephadex filtration using the SpermprepI method significantly improves sperm motility and grade in mice. Filtered sperm also show enhanced viability and longevity during in vitro incubation.
Area of Science:
- Reproductive Biology
- Sperm Sorting Technologies
- In Vitro Fertilization
Background:
- Mouse epididymal spermatozoa are crucial for reproductive studies.
- Sperm preparation methods aim to isolate motile and morphologically normal sperm.
- Assessing sperm viability and motility post-processing is essential for ART.
Purpose of the Study:
- To evaluate the efficacy of Sephadex filtration (SpermprepI) for separating motile, morphologically normal mouse epididymal spermatozoa.
- To assess the viability and motility of recovered spermatozoa over a 3-hour incubation period.
Main Methods:
- Spermatozoa were harvested from mouse epididymides and incubated in Test-Yolk buffer (TYB).
- Samples were divided into control and filtered aliquots using the SpermprepI column.
- Filtration fractions were assessed for motility and morphology, and incubated for 3 hours.
Main Results:
- SpermprepI filtration significantly improved sperm motility (91.5% vs 76.5%) and motility grade (3.0 vs 2.5).
- Filtration did not significantly alter the percentage of normal sperm morphology.
- Filtered sperm exhibited superior motility (63% vs 39%) and motility grade (1.66 vs 0.82) after 3-hour incubation compared to controls.
Conclusions:
- The SpermprepI filtration method effectively enhances sperm motility and grade.
- Filtered spermatozoa demonstrate improved in vitro longevity and motility.
- This technique selects for higher quality spermatozoa with better post-incubation performance.
