Keywords
sperm selection
microfluidic device
How to Cite
Abstract
Study question: Which sperm selection technique will most significantly reduce sperm DNA fragmentation levels?
Summary answer: The ZyMöt microfluidic system achieved the greatest reduction in sperm DNA fragmentation compared to Swim-up and Density Gradient centrifugation.
What is already known: High levels of sperm DNA fragmentation can affect reproductive success in various ways: poor embryonic quality, decreased implantation and pregnancy rates and increased conception time.
Study design: Two prospective experiments, with 25 and 19 patients respectively, conducted between January 2019 and March 2021.
Materials and Methods:Sperm DNA fragmentation was evaluatedin Experiment 1 after processing semen samples using Density Gradient centrifugation and Swim-up techniques and in Experiment 2 it was evaluated after processing semen samples using Swim-up techniques and the ZyMöt microfluidic system. To evaluate sperm DNA fragmentation post-processing Tunel technique was used. The results were analyzed using the Kruskal-Wallist and Mann Whitney tests as appropriate, considering p values <0.05 significant.
Results: In Experiment 1, the Swim-up technique showed a significantly greater decrease in sperm DNA fragmentation when compared to Density Gradient centrifugation (9.1% ± 1.9 vs. 19.2% ± 5.1). In Experiment 2, the microfluidic system showed a significantly greater decrease in sperm DNA fragmentation when compared to swim-up (1.3% ± 0.7 vs. 9.1% ± 2.0).
Wider implications of the findings: It is necessary to evaluate the usefulness of a certain method of sperm selection according to the assisted reproduction techniquewhere the enriched sample will be used. ZyMöt proved to be more effective in reducing sperm DNA fragmentation.
References
Agarwal A, Majzoub A, Baskaran S, PannerSelvam M K, Cho CL, Henkel R, Finelli R, Leisegang K, Sengupta P, Barbarosie C, ParekhN, Alves MG, Ko E, Arafa M, Tadros N, Ramasamy R, Kavoussi P, Ambar R, Kuchakulla M, Robert KA, Shah R. Sperm DNA fragmentation: A new guidelineforclinicians. TheWorldJournal of Men'sHealth. 2020; 38:412–471.
Martínez E, Bezazián C, Bezazián A, LindlK, Peliquero A, Cattaneo A, Gnocchi D, Irigoyen M, Tessari L, Martínez AG. Sperm DNA fragmentation and male age: re-sults of in vitro fertilization treatments. JBRA Assist Reprod. 2020; Nov 5. doi: 10.5935/1518-0557.20210018. Epub ahead of print. PMID: 34061484.
Ribas-Maynou J, Yeste M, Salas-Huetos A. The relationship between sperm oxida-tive stress alterations and IVF/ICSI outcomes: A systematic review from nonhuman mammals. Biology. 2020; 9:178.
Agarwal A, PannerSelvam MK, Baskaran S, Cho CL. Sperm DNA damage and its impact on male reproductive health: a critical review for clinicians, reproductive professionals and researchers. ExpertReview of Molecular Diagnostics. 2019; 19:443–457.
Aitken RJ. Impact of oxidative stress on male and female germ cells: implications for fertility. Reproduction. 2020; 159:189-201.
Chianese R, Pierantoni R. Mitochondrial reactive oxygen species (ROS) production alters sperm quality. Antioxidants. 2021; 10:92.
Asghar W, Velasco V, Kingsley JL, Shoukat MS, Shafiee H, Anchan RM, Mutter GL, Tüzel E, Demirci U. Selection of functional human sperm with higher DNA integrity and fewer reactive oxygen species. AdvancedHealthcareMaterials. 2014; 3:1671-1679.
Rappa KL, Rodriguez HF, Hakkarainen GC, Anchan RM, Mutter GL, Asghar W. Sperm processing for advanced reproductive technologies: Where are we today? BiotechnologyAdvances. 2016; 34:578–587.
Sequeira RC, Criswell T, Atala A, Yoo JJ. Microfluidic systems for assisted reproduc-tive technologies: advantages and potential applications. TissueEngineering and Regenerative Medicine. 2020; 17:787-800.
Vaughan DA, Sakkas D. Sperm selection methods in the 21st century. Biology of Reproduction, 2019; 101:1076-1082.
Gode F, Gürbüz AS, Tamer B, Pala I, Isik AZ. The effects of microfluidic sperm sort-ing, density gradient and swim-up methods on semen oxidation reduction potential. UrologyJournal. 2020; 17:397-401.
Parrella A, Keating D, Cheung S, Xie P, Stewart JD, Rosenwaks Z, Palermo GD. A treatment approach for couples with disrupted sperm DNA integrity and recurrent ART failure. Journal of AssistedReproduction and Genetics. 2019; 36:2057-2066.
WHO World Health Organization. WHO Laboratory Manual for the Examination and Processing of Human Semen, 5th edn. Geneva: WorldHealthOrganization. 2010.
Muthigi A, Jahandideh S, Bishop LA, Naeemi FK, Shipley SK, O'Brien JE, Shin PR, Devine K, Tanrikut C. Clarifying the relationship between total motile sperm counts and intrauterine insemination pregnancy rates. Fertility and Sterility. 2021; 115:1454-1460.
Nosrati R, Graham PJ, Zhang B, Riordon J, Lagunov A, Hannam TG, Escobedo C, Jarvi K, Sinton D. Microfluidicsforspermanalysis and selection. NatureReviews. Uro-logy. 2017; 14:707-730.
Samuel R, Feng H, Jafek A, Despain D, Jenkins T, Gale B. Microfluidic-based sperm sorting analysis for treatment of male infertility. TranslationalAndrology and Uro-logy. 2018; 7:336-347.
Shirota K, Yotsumoto F, Itoh H, Obama H, Hidaka N, Nakajima K, Miyamoto S. Sepa-ration efficiency of a microfluidic sperm sorter to minimize sperm DNA damage. Fertility and Sterility. 2016; 105:315–21.e1.
De Gheselle S, Deroose A, Stevens J, Hiel M, Tilleman K. A methodological valida-tion of an easy one-step swimout semen preparation procedure for selecting DNA fragmentation-free spermatozoa for ICSI. Andrologia. 2020; 52:e13852.