Comparison of In-House Microfluidic Device and Centrifuge-Based Method Efficacy in Sperm Preparation for Assisted Reproductive Technology

Phairatana, Tonghathai; Prateepchaikul, Thanaporn; Navakanittworakul, Raphatphorn; Choksuchat, Chainarong

doi:10.18502/jri.v24i2.12492

Comparison of In-House Microfluidic Device and Centrifuge-Based Method Efficacy in Sperm Preparation for Assisted Reproductive Technology

Authors

Tonghathai Phairatana ¹

Thanaporn Prateepchaikul ²

Raphatphorn Navakanittworakul ¹

Chainarong Choksuchat ²

¹ Department of Biomedical Sciences and Biomedical Engineering, Faculty of Medicine, Prince of Songkla University, Songkhla, Thailand

² Department of Obstetrics and Gynecology, Faculty of Medicine, Prince of Songkla University, Songkhla, Thailand

10.18502/jri.v24i2.12492

Abstract

Background: Sperm DNA fragmentation can affect reproductive outcomes in assisted reproductive techniques (ART), and it is a concern in density gradient centrifugation (DGC). By contrast, microfluidic approaches allow the selection of highly motile sperm with low DNA fragmentation index (DFI). The purpose of current study, was to compare the efficacy of a microfluidic device designed in-house in comparison with DGC. Methods: Nineteen healthy men with normal semen profiles were included in the study. Semen samples were individually aliquoted for three sperm preparation analyses (crude and processed with to either DGC or the microfluidic method). Sperm parameters of the samples were evaluated along with DNA fragmentation using the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) method. Results: Sperm processed using the microfluidic method showed a significantly lower DFI than those obtained using DGC and in crude semen, with DFI of 1.1%, 3.5%, and 4.9%, respectively. Although the microfluidic method yielded significantly lower sperm concentrations than DGC, no significant differences were observed in total motility, progressive motility, curvilinear velocity, straight-line velocity, or normal morphology. Conclusion: Using the in-house microfluidic device, sperm with lower DFI was effectively isolated when compared with DGC. The motility and normal morphology rates were comparable among the samples.

Keywords

Assisted Reproductive Technology

Density gradient centrifugation

DNA Fragmentation

microfluidics

Sperm

Zegers-Hochschild F, Adamson GD, Dyer S, Racowsky C, de Mouzon J, Sokol R, et al. The international glossary on infertility and fertility care, 2017. Fertil Steril. 2017;108(3):393-406.

Cui W. Mother or nothing: the agony of infertility. Bull World Health Organ. 2010;88(12):881-2.

Taylor HS, Pal L, Seli E. Speroff’s clinical gynecologic endocrinology and infertility. 9th ed. Philadelphia: Lippincott Williams & Wilkins; 2019. 1292 p.

Oseguera-López I, Ruiz-Díaz S, Ramos-Ibeas P, Pérez-Cerezales S. Novel techniques of sperm selection for improving IVF and ICSI outcomes. Front Cell Dev Biol. 2019;7:298.

Agarwal A, Baskaran S, Parekh N, Cho CL, Henkel R, Vij S, et al. Male infertility. Lancet. 2021;397 (10271):319-33.

Agarwal A, Said TM, Bedaiwy MA, Banerjee J, Alvarez JG. Oxidative stress in an assisted reproductive techniques setting. Fertil Steril. 2006;86(3): 503-12.

Ahelik A, Mändar R, Korrovits P, Karits P, Talving E, Rosenstein K, et al. Systemic oxidative stress could predict assisted reproductive technique outcome. J Assist Reprod Genet. 2015;32(5):699-704.

Muriel L, Garrido N, Fernández JL, Remohí J, Pellicer A, de los Santos MJ, et al. Value of the sperm deoxyribonucleic acid fragmentation level, as measured by the sperm chromatin dispersion test, in the outcome of in vitro fertilization and intracytoplasmic sperm injection. Fertil Steril. 2006;85(2):371-83.

Velez de la Calle JF, Muller A, Walschaerts M, Clavere JL, Jimenez C, Wittemer C, et al. Sperm deoxyribonucleic acid fragmentation as assessed by the sperm chromatin dispersion test in assisted reproductive technology programs: results of a large prospective multicenter study. Fertil Steril. 2008; 90(5):1792-9.

Borini A, Tarozzi N, Bizzaro D, Bonu MA, Fava L, Flamigni C, et al. Sperm DNA fragmentation: paternal effect on early post-implantation embryo development in ART. Hum Reprod. 2006;21(11): 2876-81.

Jerre E, Bungum M, Evenson D, Giwercman A. Sperm chromatin structure assay high DNA stainability sperm as a marker of early miscarriage after intracytoplasmic sperm injection. Fertil Steril. 2019;112(1):46-53.e2.

Nicopoullos J, Vicens-Morton A, Lewis SEM, Lee K, Larsen P, Ramsay J, et al. Novel use of COMET parameters of sperm DNA damage may increase its utility to diagnose male infertility and predict live births following both IVF and ICSI. Hum Reprod. 2019;34(10):1915-23.

Robinson L, Gallos ID, Conner SJ, Rajkhowa M, Miller D, Lewis S, et al. The effect of sperm DNA fragmentation on miscarriage rates: a systematic review and meta-analysis. Hum Reprod. 2012;27(10):2908-17.

Zhao J, Zhang Q, Wang Y, Li Y. Whether sperm deoxyribonucleic acid fragmentation has an effect on pregnancy and miscarriage after in vitro fertilization/intracytoplasmic sperm injection: a systematic review and meta-analysis. Fertil Steril. 2014;102(4):998-1005.e8.

Chinnasamy T, Kingsley JL, Inci F, Turek PJ, Rosen MP, Behr B, et al. Guidance and self-sorting of active swimmers: 3D periodic arrays increase persistence length of human sperm selecting for the fittest. Adv Sci (Weinh). 2017;5(2):1700531.

Gode F, Gürbüz AS, Tamer B, Pala I, Isik AZ. The effects of microfluidic sperm sorting, density gradient and swim-up methods on semen oxidation reduction potential. Urol J. 2020;17(4):397-401.

Kishi K, Ogata H, Ogata S, Mizusawa Y, Okamoto E, Matsumoto Y, et al. Frequency of sperm DNA fragmentation according to selection method: comparison and relevance of a microfluidic device and a swim-up procedure. J Clin Diagn Res. 2015;9(11):QC14-6.

Asghar W, Velasco V, Kingsley JL, Shoukat MS, Shafiee H, Anchan RM, et al. Selection of functional human sperm with higher DNA integrity and fewer reactive oxygen species. Adv Healthc Mater. 2014;3(10):1671-9.

Quinn MM, Jalalian L, Ribeiro S, Ona K, Demirci U, Cedars MI, et al. Microfluidic sorting selects sperm for clinical use with reduced DNA damage compared to density gradient centrifugation with swim-up in split semen samples. Hum Reprod. 2018;33(8):1388-93.

Shirota K, Yotsumoto F, Itoh H, Obama H, Hidaka N, Nakajima K, et al. Separation efficiency of a microfluidic sperm sorter to minimize sperm DNA damage. Fertil Steril. 2016;105(2):315-21.e1.

World Health Organization. WHO Laboratory manual for the examination and processing of human semen. 5th ed. Geneva: World Health Organization; 2010. 286 p.

Wei-Xuan LI, Liang GT, Wei YA, Zhang Q, Wei WA, Xiao-Mian ZH, et al. Artificial uterus on a microfluidic chip. Chinese J Analytical Chem. 2013;41(4):467-72.

Mancini V, Pensabene V. Organs-on-chip models of the female reproductive system. Bioengineering (Basel). 2019;6(4):103.

Ahmadkhani N, Hosseini M, Saadatmand M, Abbaspourrad A. The influence of the female reproductive tract and sperm features on the design of microfluidic sperm-sorting devices. J Assist Reprod Genet. 2022;39(1):19-36.

Barroso G, Morshedi M, Oehninger S. Analysis of DNA fragmentation, plasma membrane translocation of phosphatidylserine and oxidative stress in human spermatozoa. Hum Reprod. 2000;15(6): 1338-44.

Muratori M, Tarozzi N, Carpentiero F, Danti S, Perrone FM, Cambi M, et al. Sperm selection with density gradient centrifugation and swim up: effect on DNA fragmentation in viable spermatozoa. Sci Rep. 2019;9(1):7492.

Xue LT, Wang RX, He B, Mo WY, Huang L, Wang SK, et al. Effect of sperm DNA fragmentation on clinical outcomes for Chinese couples undergoing in vitro fertilization or intracytoplasmic sperm injection. J Int Med Res. 2016;44(6):1283-91.

Ni W, Xiao S, Qiu X, Jin J, Pan C, Li Y, et al. Effect of sperm DNA fragmentation on clinical outcome of frozen–thawed embryo transfer and on blastocyst formation. PLoS One. 2014;9(4): e94956.

Sedo CA, Bilinski M, Lorenzi D, Uriondo H, Noblía F, Longobucco V, et al. Effect of sperm DNA fragmentation on embryo development: clinical and biological aspects. JBRA Assist Reprod. 2017;21(4):343-50.

Jin J, Pan C, Fei Q, Ni W, Yang X, Zhang L, et al. Effect of sperm DNA fragmentation on the clinical outcomes for in vitro fertilization and intracytoplasmic sperm injection in women with different ovarian reserves. Fertil Steril. 2015;103(4):910-6.

Lin MH, Lee RKK, Li SH, Lu CH, Sun FJ, Hwu YM. Sperm chromatin structure assay parameters are not related to fertilization rates, embryo quality, and pregnancy rates in in vitro fertilization and intracytoplasmic sperm injection, but might be related to spontaneous abortion rates. Fertil Steril. 2008;90(2):352-9.