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Home / Equine Research Bank / Theriogenology / High pressure flow cytometric sorting damages sperm.
Theriogenology2005; 64(5); 1035-1048; doi: 10.1016/j.theriogenology.2005.02.002

High pressure flow cytometric sorting damages sperm.

Abstract: Sexing sperm by high-speed flow cytometry subjects them to high pressure. The routine operating pressure of the MoFlo SX flow cytometer for sperm sorting for commercial production has been 50 pounds/square inch (psi), with a standard 70 microm standard nozzle tip. It was hypothesized that lowering the sorting pressure could reduce sperm damage. Therefore, a series of experiments using semen from six bulls, sorted with three MoFlo SX sorters, was conducted to determine optimal pressure. An additional experiment was done with stallion spermatozoa. In Experiment 1, sorting at 30 psi compared to 50 psi with the 70 microm nozzle tip increased sperm motility post-thaw at 30 min and 2h from 40.5 to 48.0% and 30.0 to 40.2%, respectively (P<0.05). In Experiment 2, 49, 43, 37, 31, and 25 psi resulted in 24.2, 32.8, 35.6, 37.5, and 39.8% progressively motile spermatozoa post-thaw (P<0.05). In Experiment 3, 3 pressures (50, 40, 30 psi)x2 sorting methods were further evaluated. At 50, 40, and 30 psi, respective mean sperm motilities at 30 min were 44.8, 48.6, and 49.6% (P<0.05), and percentage of live spermatozoa were 51.7, 55.7, and 57.8% (P<0.05). The improvement of post-sort sperm quality with lowered pressure was also evident in stallion spermatozoa. After sorting at 30, 40 and 50 psi were 40.6, 34.5 and 30.1% motile spermatozoa (P<0.1), and were 76.7, 72.5 and 67.8% (P0.05), and higher sperm concentration during staining increased sort rate (P<0.05). In conclusion, lowering pressure of the MoFlo SX flow cytometer for sperm sorting from 50 psi (standard pressure) to 40 psi clearly improved sperm quality without a significant decrease in sorter performance.
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Publication Date: 2005-03-02 PubMed ID: 16125550DOI: 10.1016/j.theriogenology.2005.02.002Google Scholar: Lookup
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Summary

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This paper investigates the impact of high-pressure conditions during sperm sorting using the MoFlo SX flow cytometer on the quality of sperm. The research found that reducing the sorting pressure from the standard measure of 50 psi improves sperm quality without affecting the cytometer’s performance.

Article Explanation

This research performed a series of experiments to ascertain whether reducing the pressures used in a MoFlo SX flow cytometer for sperm sorting could result in minimized sperm damage. This is critical because the cytometer’s standard operating pressure was 50 psi, which could result in a substantial degradation of sperm quality.

  • In the first experiment, lowering the pressure to 30 psi from 50 psi with a 70 micrometre nozzle tip led to a considerable increase in sperm motility post-thaw at both 30 minutes and 2 hours, demonstrating that pressure reduction is beneficial for sperm quality.

Further Findings

The study did not stop at the first experiment and conducted further tests to solidify its findings.

  • The second experiment showed that further reducing the pressure (to 49, 43, 37, 31, and 25 psi) led to a progressive increase in the percentage of motile sperm post-thaw, affirming the notion that lower pressure yields superior sperm quality.
  • The third experiment analyzed three different pressure levels combined with two sorting methods. At each respective pressure level (50, 40, and 30 psi), the mean sperm motilities increased, as did the percentage of live spermatozoa; therefore, further pressurizing the evidence of lower pressure resulting in increased sperm quality.
  • The same observations were made when the researchers carried out the same experiment in stallion spermatozoa. Lower pressure seemed to yield superior sperm quality in these animals.

Impact on Cytometer Performance

Aside from assessing sperm quality, the study also took into account the cytometer’s performance at different pressure levels.

  • In the fourth experiment, two pressures (40 and 50 psi) and two staining concentrations of bovine spermatozoa were evaluated. Notably, the research found that lowering the pressure to 40 psi did not affect the sort rate and purity of the sperm. Furthermore, increasing the sperm concentration during staining led to a higher sort rate.
  • It was concluded that reducing the MoFlo SX flow cytometer’s sperm sorting pressure from 50 psi to 40 psi improves sperm quality without significantly affecting the cytometer’s performance.

Cite This Article

APA
Suh TK, Schenk JL, Seidel GE. (2005). High pressure flow cytometric sorting damages sperm. Theriogenology, 64(5), 1035-1048. https://doi.org/10.1016/j.theriogenology.2005.02.002

Publication

Theriogenology
ISSN: 0093-691X
NlmUniqueID: 0421510
Country: United States
Language: English
Volume: 64
Issue: 5
Pages: 1035-1048

Researcher Affiliations

Suh, T K
  • Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523, USA. tksuh@colostate.edu
Schenk, J L
    Seidel, G E

      MeSH Terms

      • Animals
      • Cattle
      • Cell Separation
      • Cell Survival
      • Flow Cytometry / veterinary
      • Horses
      • Male
      • Pressure / adverse effects
      • Sex Determination Analysis / veterinary
      • Sperm Count
      • Sperm Motility
      • Spermatozoa / physiology

      Citations

      This article has been cited 17 times.
      1. Uhm SJ, Heo YT, Yu DM, Kim DK, Gupta MK. Pre-implantation development of cattle embryos produced from fresh bull semen enriched for X- chromosome-bearing spermatozoa using a monoclonal antibody.. Vet Res Commun 2023 Jul 11;.
        doi: 10.1007/s11259-023-10160-8pubmed: 37432602google scholar: lookup
      2. Box A, Holmes L, DeLay M, Adams D, Bergeron A, Clise-Dwyer K, Cochran M, Guerra RDR, Meyer EM, Brundage KM. Cell Sorter Cleaning Practices and Their Impact on Instrument Sterility.. J Biomol Tech 2022 Apr 15;33(1).
        doi: 10.7171/3fc1f5fe.e2675d74pubmed: 35837003google scholar: lookup
      3. Moradi M, Hajarian H, Karamishabankareh H, Soltani L, Soleymani B. Pre-treatment of ram semen extender with magnetic nanoparticles on freeze-thawed spermatozoa.. Vet Med Sci 2022 Mar;8(2):792-798.
        doi: 10.1002/vms3.689pubmed: 34914199google scholar: lookup
      4. Orsolini MF, Meyers SA, Dini P. An Update on Semen Physiology, Technologies, and Selection Techniques for the Advancement of In Vitro Equine Embryo Production: Section II.. Animals (Basel) 2021 Nov 20;11(11).
        doi: 10.3390/ani11113319pubmed: 34828049google scholar: lookup
      5. Chen PR, Redel BK, Kerns KC, Spate LD, Prather RS. Challenges and Considerations during In Vitro Production of Porcine Embryos.. Cells 2021 Oct 15;10(10).
        doi: 10.3390/cells10102770pubmed: 34685749google scholar: lookup
      6. He Q, Wu S, Huang M, Wang Y, Zhang K, Kang J, Zhang Y, Quan F. Effects of Diluent pH on Enrichment and Performance of Dairy Goat X/Y Sperm.. Front Cell Dev Biol 2021;9:747722.
        doi: 10.3389/fcell.2021.747722pubmed: 34660605google scholar: lookup
      7. Reese S, Pirez MC, Steele H, Kölle S. The reproductive success of bovine sperm after sex-sorting: a meta-analysis.. Sci Rep 2021 Aug 30;11(1):17366.
        doi: 10.1038/s41598-021-96834-2pubmed: 34462506google scholar: lookup
      8. Keles E, Malama E, Bozukova S, Siuda M, Wyck S, Witschi U, Bauersachs S, Bollwein H. The micro-RNA content of unsorted cryopreserved bovine sperm and its relation to the fertility of sperm after sex-sorting.. BMC Genomics 2021 Jan 7;22(1):30.
        doi: 10.1186/s12864-020-07280-9pubmed: 33413071google scholar: lookup
      9. Xie Y, Xu Z, Wu Z, Hong L. Sex Manipulation Technologies Progress in Livestock: A Review.. Front Vet Sci 2020;7:481.
        doi: 10.3389/fvets.2020.00481pubmed: 32923466google scholar: lookup
      10. Hamacher T, Berendsen JTW, Kruit SA, Broekhuijse MLWJ, Segerink LI. Effect of microfluidic processing on the viability of boar and bull spermatozoa.. Biomicrofluidics 2020 Jul;14(4):044111.
        doi: 10.1063/5.0013919pubmed: 32774586google scholar: lookup
      11. Steele H, Makri D, Maalouf WE, Reese S, Kölle S. Bovine Sperm Sexing Alters Sperm Morphokinetics and Subsequent Early Embryonic Development.. Sci Rep 2020 Apr 10;10(1):6255.
        doi: 10.1038/s41598-020-63077-6pubmed: 32277124google scholar: lookup
      12. Khoramian Tusi B, Socolovsky M. High-throughput single-cell fate potential assay of murine hematopoietic progenitors in vitro.. Exp Hematol 2018 Apr;60:21-29.e3.
        doi: 10.1016/j.exphem.2018.01.005pubmed: 29410050google scholar: lookup
      13. Liu L, Nielsen FM, Riis SE, Emmersen J, Fink T, Hjortdal JØ, Bath C, Zachar V. Maintaining RNA Integrity for Transcriptomic Profiling of Ex Vivo Cultured Limbal Epithelial Stem Cells after Fluorescence-Activated Cell Sorting (FACS).. Biol Proced Online 2017;19:15.
        doi: 10.1186/s12575-017-0065-2pubmed: 29255379google scholar: lookup
      14. Shields CW 4th, Ohiri KA, Szott LM, López GP. Translating microfluidics: Cell separation technologies and their barriers to commercialization.. Cytometry B Clin Cytom 2017 Mar;92(2):115-125.
        doi: 10.1002/cyto.b.21388pubmed: 27282966google scholar: lookup
      15. Chen TJ, Wu JK, Chang YC, Fu CY, Wang TP, Lin CY, Chang HY, Chieng CC, Tzeng CY, Tseng FG. High-efficiency rare cell identification on a high-density self-assembled cell arrangement chip.. Biomicrofluidics 2014 May;8(3):036501.
        doi: 10.1063/1.4874716pubmed: 24926391google scholar: lookup
      16. Barteneva NS, Ponomarev ED, Tsytsykova A, Armant M, Vorobjev IA. Mitochondrial staining allows robust elimination of apoptotic and damaged cells during cell sorting.. J Histochem Cytochem 2014 Apr;62(4):265-75.
        doi: 10.1369/0022155413520404pubmed: 24394470google scholar: lookup
      17. Espinosa-Cervantes R, Córdova-Izquierdo A. Sexing sperm of domestic animals.. Trop Anim Health Prod 2013 Jan;45(1):1-8.
        doi: 10.1007/s11250-012-0215-0pubmed: 22829354google scholar: lookup
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