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Methods in molecular biology (Clifton, N.J.)2012; 927; 279-295; doi: 10.1007/978-1-62703-038-0_26

Sex-sorting sperm using flow cytometry/cell sorting.

Abstract: The sex of mammalian offspring can be predetermined by flow sorting relatively pure living populations of X- and Y-chromosome-bearing sperm. This method is based on precise staining of the DNA of sperm with the nucleic acid-specific fluorophore, Hoechst 33342, to differentiate between the subpopulations of X- and Y-sperm. The fluorescently stained sperm are then sex-sorted using a specialized high speed sorter, MoFlo(®) SX XDP, and collected into biologically supportive media prior to reconcentration and cryopreservation in numbers adequate for use with artificial insemination for some species or for in vitro fertilization. Sperm sorting can provide subpopulations of X- or Y-bearing bovine sperm at rates in the 8,000 sperm/s range while maintaining; a purity of 90% such that it has been applied to cattle on a commercial basis. The sex of offspring has been predetermined in a wide variety of mammalian species including cattle, swine, horses, sheep, goats, dogs, cats, deer, elk, dolphins, water buffalo as well as in humans using flow cytometric sorting of X- and Y-sperm.
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Publication Date: 2012-09-21 PubMed ID: 22992923DOI: 10.1007/978-1-62703-038-0_26Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research studied a method for pre-determining the sex of mammalian offspring by sorting sperm with the help of flow cytometry. This technique allows scientists to separate and collect X-chromosome and Y-chromosome bearing sperm that have been identified with a DNA-specific stain, and has been used successfully in numerous species, including humans and farm animals.

Overview of the Research

  • The goal of this research was to verify whether or not the sex of mammalian offspring could be predetermined by using flow cytometry/cell sorting to separate relatively pure populations of X- and Y-chromosome-bearing sperm.
  • The team employed a specialized method which involved staining sperm DNA with the nucleic acid-specific fluorophore, Hoechst 33342, which is capable of differentiating between X-bearing and Y-bearing sperm due to differing DNA content.

The Methodology

  • Once the sperm had been stained using the fluorescent compound Hoechst 33342, they were sorted using a high-speed cell sorter, the MoFlo(®) SX XDP.
  • Following sorting, the sperm were transferred into a biologically supportive medium in preparation for reconcentration and cryopreservation.
  • The sorting process was designed to gather sperm in quantities suitable for use in artificial insemination procedures for certain species, or for in vitro fertilization.

Results and Implications

  • The researchers found that the flow cytometric sorting procedure could isolate X- or Y-bearing sperm in the 8,000 sperm/second range while maintaining an approximate purity of 90%.
  • This level of success enabled the technique to be applied on a commercial basis for cattle, but it proved effective across various mammalian species including swine, horses, sheep, goats, dogs, cats, deer, elk, dolphins, water buffalo and even humans.
  • The ability to predetermine the sex of offspring using this method potentially allows for greater control in breeding programs, whether they be aimed at livestock farming, preservation of endangered species or human reproductive therapies.

Cite This Article

APA
Garner DL, Evans KM, Seidel GE. (2012). Sex-sorting sperm using flow cytometry/cell sorting. Methods Mol Biol, 927, 279-295. https://doi.org/10.1007/978-1-62703-038-0_26

Publication

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
NlmUniqueID: 9214969
Country: United States
Language: English
Volume: 927
Pages: 279-295

Researcher Affiliations

Garner, Duane L
  • GametoBiology Consulting, Graeagle, CA, USA. duanejunegarner@msn.com
Evans, K Michael
    Seidel, George E

      MeSH Terms

      • Animals
      • Cell Separation / instrumentation
      • Cell Separation / methods
      • Flow Cytometry / instrumentation
      • Flow Cytometry / methods
      • Humans
      • Male
      • Sex Preselection / methods
      • Spermatozoa / cytology
      • Spermatozoa / metabolism
      • X Chromosome / metabolism
      • Y Chromosome / metabolism

      Citations

      This article has been cited 16 times.
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