Improvement of bovine semen quality by removal of membrane-damaged sperm cells with DNA aptamers and magnetic nanoparticles

FARINI, VERONICA L.; CAMAÑO, CARLA V.; YBARRA, GABRIEL; VIALE, DIEGO L.; VICHERA, GABRIEL; YAKISICH, JUAN S.; RADRIZZANI, MARTÍN

JOURNAL OF BIOTECHNOLOGY

ELSEVIER SCIENCE BV

Año: 2016 vol. 229 p. 33 - 41

0168-1656

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 thequality of semen prior to AI. In thiswork we used the Cell-SELEX technique to select single strand DNA aptamers ableto recognize with high affinity and specificity damaged sperm cells generatedby heat-treatment. We first isolated aptamers with a conserved two motifs of 6nucleotides of length that bind to the membrane of heat-treated spermatozoa.Then, we used synthetic biotin-labeled aptamers containing the conserved motifto recognize membrane-damaged cells and separate them from viable cells by theuse of avidin-coated superparamagnetic iron oxide nanoparticles (SPION). Thisprocedure improved the quality of semen by significantly increasing thepercentage of healthy sperm cells without affecting the rate of blastocystcleavage. This technique was successfullyapplied to both unsorted and sex-sorted spermsuspension.