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.
Publication Date: 2012-09-21 PubMed ID: 22992923DOI: 10.1007/978-1-62703-038-0_26Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Review
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
Researcher Affiliations
- GametoBiology Consulting, Graeagle, CA, USA. duanejunegarner@msn.com
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.- Meunier L, Sorci G, Silva Vieira C, Sadiq Shiek S, Hingrat Y, Saint Jalme M, Torres Carreira J. Exploring the mechanisms underlying the last male precedence in the North African houbara bustard. J Exp Biol 2025 Dec 15;228(24).
- Chen H, Rushdi HE, Loor JJ, Teng Z, Liu S. Progress of sex control techniques in mammals†. Biol Reprod 2025 Nov 14;113(5):997-1012.
- Bai L, Zhao Y, Zhou Y, Song Y, Xiao H, Zhao G, Wang Z, Li X. Advances in immunological sorting of X and Y chromosome-bearing sperm: from proteome to sex-specific proteins. Front Vet Sci 2025;12:1523491.
- Singh SK, Kumar R, Mathur M, Kamboj H, Kaushik JK, Mohanty AK, Kumar S. Exploring aptamers for targeted enrichment of X sperm in bovine: unraveling selective potential. Anim Biotechnol 2024 Nov;35(1):2323592.
- Pinto-Pinho P, Ferreira AF, Pinto-Leite R, Fardilha M, Colaço B. The History and Prospects of Rabbit Sperm Sexing. Vet Sci 2023 Aug 7;10(8).
- Yotov S, Fasulkov I, Atanasov A, Kistanova E, Sinapov B, Ivanova B, Yarkov D, Zaimova D. Influence of Ovarian Status and Steroid Hormone Concentration on Day of Timed Artificial Insemination (TAI) on the Reproductive Performance of Dairy Cows Inseminated with Sexed Semen. Animals (Basel) 2023 Mar 1;13(5).
- He Q, Gao F, Wu S, Wang S, Xu Z, Xu X, Lan T, Zhang K, Quan F. Alkaline Dilution Alters Sperm Motility in Dairy Goat by Affecting sAC/cAMP/PKA Pathway Activity. Int J Mol Sci 2023 Jan 16;24(2).
- Hu C, Shi J, Chi Y, Yang J, Cui Q. Y/X-Chromosome-Bearing Sperm Shows Elevated Ratio in the Left but Not the Right Testes in Healthy Mice. Life (Basel) 2021 Nov 11;11(11).
- 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).
- 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.
- Quelhas J, Santiago J, Matos B, Rocha A, Lopes G, Fardilha M. Bovine semen sexing: Sperm membrane proteomics as candidates for immunological selection of X- and Y-chromosome-bearing sperm. Vet Med Sci 2021 Sep;7(5):1633-1641.
- Xie Y, Xu Z, Wu Z, Hong L. Sex Manipulation Technologies Progress in Livestock: A Review. Front Vet Sci 2020;7:481.
- Bechsgaard J, Schou MF, Vanthournout B, Hendrickx F, Knudsen B, Settepani V, Schierup MH, Bilde T. Evidence for Faster X Chromosome Evolution in Spiders. Mol Biol Evol 2019 Jun 1;36(6):1281-1293.
- Naniwa Y, Sakamoto Y, Toda S, Uchiyama K. Bovine sperm sex-selection technology in Japan. Reprod Med Biol 2019 Jan;18(1):17-26.
- Khamlor T, Pongpiachan P, Sangsritavong S, Chokesajjawatee N. Determination of Sperm Sex Ratio in Bovine Semen Using Multiplex Real-time Polymerase Chain Reaction. Asian-Australas J Anim Sci 2014 Oct;27(10):1411-6.
- Dezhkam L, Dezhkam H, Dezhkam I. Sex selection from Islamic point of view. Iran J Reprod Med 2014 Apr;12(4):289-90.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
