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- Title
Magnetic activated cell sorting and its application for selection of human non apoptotic spermatozoa in ART.
- Authors
Hassani Bafrani H.; Catt S.; Pangestu M.; Smith Peter T.
- Abstract
Introduction: Flydrogen peroxide has been implemented in literature to induce a significant increase in caspase. Activation of caspase 9 triggers a cascade of caspase activation, including caspase 3, which promotes cellular apoptosis. Magnetic cell sorting (MACS) using annexin V-conjugated microbeads eliminates apoptotic spermatozoa with annexin V-positive. Materials and Methods: A total of 20 semen samples were obtained from male partners of couples for analysis. One aliquot (0.5ml) of the sperm suspension was subjected to MACS. Motility and concentration was checked, and a sample taken for tunnel before and after MACS. The remaining sperm suspension was divided into 6 tubes (2x control, 2x peroxide, 2x peroxide/ melatonin). DNA fragmentation was evaluated using the TUNEL assay (Roche, Indianapolis, IN, USA) (14), with some modifications. Data were analyzed using Graph Pad Instat Ver. 3.10. Results: Results of the TUNEL assay in pretreatments of human spermatozoa with 1 OOuM peroxidase for 24 hrs revealed that the percentage of sperm with fragmented DNA was significantly lower on the sored sperm after sorting MACS (p<0.001 vs. control). Following the pretreated human spermatozoa with peroxidase for 24 hr, percentage of sperm motility and progressive motility were significantly reduced (p<0.001 vs. control). Pretreatments of human spermatozoa for 24 hr revealed that the percentage of sperm motility and progressive motility were significantly reduced by 100 pM peroxidase and peroxidase with MACS, versus zero hour in control group (p<0.001). Conclusion: The sperm was treated with 100 pM peroxidase sorting using MACS retain appropriate spermatozoa and select sperm good quality. The use of MACS will select only sperm with intact this result in high percentage motility and progressive motility.
- Subjects
APOPTOSIS; SPERMATOZOA; CASPASES; PEROXIDES; MELATONIN
- Publication
Iranian Journal of Reproductive Medicine, 2015, p60
- ISSN
1680-6433
- Publication type
Article