title: Improvement of bovine semen quality by removal of membrane-damaged sperm cells with DNA aptamers and magnetic nanoparticles.
abstract: In cattle, cryopreservation of semen and sex-sorting kill up to 50% of spermatozoa and decrease the success of assisted insemination (AI). Therefore, significant efforts are being carried out to improve the quality of semen prior to AI. In this work we used the Cell-SELEX technique to select single strand DNA aptamers able to recognize with high affinity and specificity damaged sperm cells generated by heat-treatment. We first isolated aptamers with a conserved two motifs of 6 nucleotides of length that bind to the membrane of heat-treated spermatozoa. Then, we used synthetic biotin-labeled aptamers containing the conserved motif to recognize membrane-damaged cells and separate them from viable cells by the use of avidin-coated superparamagnetic iron oxide nanoparticles (SPION). This procedure improved the quality of semen by significantly increasing the percentage of healthy sperm cells without affecting the rate of blastocyst cleavage. This technique was successfully applied to both unsorted and sex-sorted sperm suspension.
authors.affiliation: Laboratory of Neurogenetic and Molecular Cytogenetic, School of Science and Technology, National University of San Martin, CONICET, Buenos Aires, Argentina.
authors.forename: Veronica L
authors.initials: VL
authors.lastname: Farini
authors.affiliation: Research Center on Surface Processes, Nanomaterials Unit, National Institute of Industrial Technology (INTI), Buenos Aires, Argentina.
authors.forename: Carla V
authors.initials: CV
authors.lastname: Camaño
authors.affiliation: Research Center on Surface Processes, Nanomaterials Unit, National Institute of Industrial Technology (INTI), Buenos Aires, Argentina.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Ybarra
authors.affiliation: Laboratory of Neurogenetic and Molecular Cytogenetic, School of Science and Technology, National University of San Martin, CONICET, Buenos Aires, Argentina.
authors.forename: Diego L
authors.initials: DL
authors.lastname: Viale
authors.affiliation: Laboratory of Neurogenetic and Molecular Cytogenetic, School of Science and Technology, National University of San Martin, CONICET, Buenos Aires, Argentina.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Vichera
authors.affiliation: School of Pharmacy, Department of Pharmaceutical Sciences, Hampton University, Hampton, VA, USA.
authors.forename: Juan S
authors.initials: JS
authors.lastname: Yakisich
authors.affiliation: Laboratory of Neurogenetic and Molecular Cytogenetic, School of Science and Technology, National University of San Martin, CONICET, Buenos Aires, Argentina. Electronic address: martin.radrizzani@gmail.com.
authors.forename: Martín
authors.initials: M
authors.lastname: Radrizzani
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D002417:Cattle; D002469:Cell Separation / Q000379:methods*; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D008297:Male; D052156:SELEX Aptamer Technique / Q000379:methods*; D055101:Semen Analysis / Q000379:methods*; D013094:Spermatozoa / Q000166:cytology* / Q000502:physiology
chemical_list: D052157:Aptamers, Nucleotide; D058185:Magnetite Nanoparticles
keywords: Bovine; Cell-SELEX; DNA aptamer; Magnetic nanoparticles; Semen quality