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DNA fragmentation in human sperm after magnetic-activated cell sorting
Sara Bucar1, Ana Gonçalves, Eduardo Rocha
1Department of Microscopy, Laboratory of Cell Biology, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Multidisciplinary Unit for Biomedical Research (UMIB), Rua Jorge Viterbo Ferreira, 228, 4050-313, Porto, Portugal.
Journal of Assisted Reproduction and Genetics
|November 7, 2014
Summary
Magnetic-activated cell sorting (MACS) significantly reduces sperm DNA fragmentation, especially when combined with density gradient centrifugation and swim-up techniques. This approach enhances sperm quality for assisted reproductive technologies.
Area of Science:
- Reproductive Biology
- Spermatozoa Analysis
- Assisted Reproductive Technology
Background:
- Apoptotic spermatozoa can negatively impact assisted reproductive technique (ART) success rates.
- Sperm DNA fragmentation (sDNAfrag) is a key indicator of sperm quality and potential ART outcomes.
- Magnetic-activated cell sorting (MACS) is a technique to isolate non-apoptotic spermatozoa.
Purpose of the Study:
- To evaluate the efficacy of MACS in reducing sDNAfrag.
- To compare different combinations of MACS with density gradient centrifugation (DGC) and swim-up (SU) for sDNAfrag reduction.
- To identify the optimal protocol for minimizing sDNAfrag.
Main Methods:
- Hundred semen samples were analyzed.
- Five groups were created, utilizing combinations of MACS, DGC, and SU techniques.
- Sperm DNA fragmentation was quantified using the TUNEL assay.
Main Results:
- The MACS-DGC-SU protocol achieved the highest sDNAfrag reduction rate (83.3%).
- Significant sDNAfrag reduction was observed with DGC-SU, DGC-MACS-SU, DGC-SU-MACS, and MACS-SU protocols.
- Higher sDNAfrag reduction correlated negatively with sperm vitality, membrane integrity, and progressive motility.
Conclusions:
- MACS is a promising technique for optimizing sDNAfrag reduction in ART.
- Applying MACS before DGC and SU is particularly effective.
- This method is especially beneficial for samples with low progressive motility, vitality, and hypoosmotic swelling test results.
