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Sperm Preparation by Washing and Swim-up Method Improves In Vitro Fertilization and Birth Rate in Mice

  • METHODS AND MODELS OF DEVELOPMENTAL BIOLOGY
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Abstract

The method of preparing sperm supposedly affects the success rate of in vitro fertilization. This study aimed to investigate the effect of different sperm preparation methods on in vitro fertilization and birth rate in mice. Semen obtained from two C57bl6 and NMRI strains of mice were used for sperm preparation and in vitro fertilization. Sperm preparation was done using a directly add the sperms (G1), as density gradient centrifugation method (G2), along with swim-up of the pre-washed sperm method (G3). Two-cell embryos obtained from in vitro fertilization were then transferred into pseudopregnant mice. The quality of sperm, the rate of in vitro fertilization, and the birth rate were assessed in all three study groups. Significant differences were observed in motility between the control and swim-up (NMRI mice: P = 0.008 and C57 mice: P = 0.016) and the density gradient centrifugation (DGC) and swim-up (NMRI mice: P = 0.04 and C57 mice: P = 0.02) methods. Sperm concentration was significantly higher using the control method compared to the DGC (P = 0.002) and swim-up methods (P = 0.002) in both NMRI and C57 mice groups. Sperm viability in NMRI (87.7%) and C57 (80.6%) mice was significantly higher (P = 0.001) using the swim-up method compared to the other methods. The rates of in vitro fertilization and birth have significantly increased by the swim-up method. It can be concluded that the swim-up of pre-washed sperm as an efficient and inexpensive method can be considered as an alternative for the common method of preparing sperm for in vitro fertilization in mice.

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REFERENCES

  1. Ahmadi, A., Ng, S.C., Liow, S.L., et al., Fertilization and early embryology: Intracytoplasmic sperm injection of mouse oocytes with 5 mM Ca2+ at different intervals, Hum. Reprod., 1995, vol. 10, pp. 431–435.

    Article  CAS  Google Scholar 

  2. Almeida, S., Rato, L., Sousa, M., et al., Fertility and sperm quality in the aging male, Curr. Pharm. Design, 2017, vol. 23, pp. 4429–4437.

    CAS  Google Scholar 

  3. Arias M.E., Andara K., Briones E., et al., Bovine sperm separation by Swim-up and density gradients (Percoll and BoviPure): effect on sperm quality, function and gene expression, Reprod. Biol., 2017, vol. 17, pp. 126–132.

    Article  Google Scholar 

  4. Cardona Barberán, A., Boel, A., Vanden Meerschaut, F., et al., Diagnosis and treatment of male infertility-related fertilization failure, J. Clin. Med., 2020, vol. 9, p. 3899.

    Article  Google Scholar 

  5. Chemes, H.E., Phenotypic varieties of sperm pathology: genetic abnormalities or environmental influences can result in different patterns of abnormal spermatozoa, Anim. Reprod. Sci., 2018, vol. 194, pp. 41–56.

    Article  Google Scholar 

  6. Chen, L., Li, D., Ni, X., et al., Effects of the normal sperm morphology rate on the clinical and neonatal outcomes of conventional IVF cycles, Andrologia, 2020, vol. 52, no. 5, p. 3568.

    Google Scholar 

  7. Furimsky, A., Vuong, N., Xu, H., et al., Percoll gradient-centrifuged capacitated mouse sperm have increased fertilizing ability and higher contents of sulfogalactosylglycerolipid and docosahexaenoic acid-containing phosphatidylcholine compared to washed capacitated mouse sperm, Biol. Reprod., 2005, vol. 72, pp. 574–583.

    Article  CAS  Google Scholar 

  8. Henkel, R.R. and Schill, W.-B., Sperm preparation for ART, Reprod. Biol. Endocrinol., 2003, vol. 1, p. 108.

    Article  Google Scholar 

  9. Jameel, T., Sperm swim-up: a simple and effective technique of semen processing for intrauterine insemination, JPMA, J. Pak. Med. Assoc., 2008, vol. 58, p. 71.

    Google Scholar 

  10. Martí, E., Pérez Pé, R., Muiño Blanco, T., et al., Comparative study of four different sperm washing methods using apoptotic markers in ram spermatozoa, J. Androl., 2006, vol. 27, pp. 746–753.

    Article  Google Scholar 

  11. Mochida, K., Development of assisted reproductive technologies in small animal species for their efficient preservation and production, J. Reprod. Dev., 2020.

  12. Mochida, K., Hasegawa, A., Otaka, N., et al., Devising assisted reproductive technologies for wild-derived strains of mice: 37 strains from five subspecies of Mus musculus, PLoS One, 2014, vol. 9, p. 114305.

    Article  Google Scholar 

  13. Mortimer, D., Sperm recovery techniques to maximize fertilizing capacity, Reprod., Fertil., Dev., 1994, vol. 6, pp. 25–31.

    Article  CAS  Google Scholar 

  14. Morshedi, M., Duran, H.E., Taylor, S., and Oehninger, S., Efficacy and pregnancy outcome of two methods of semen preparation for intrauterine insemination: a prospective randomized study, Fertil. Steril., 2003, vol. 79, pp. 1625–1632.

  15. Nagy, A., Gertsenstein, M., Vintersten, K., et al., Manipulating the Mouse Embryo: A Laboratory Manual, Firefly Books, 2003.

    Google Scholar 

  16. Natali, I., Sperm preparation techniques for artificial insemination-comparison of sperm washing, swim up, and density gradient centrifugation methods, in Artificial Insemination in Farm Animals, Dalam: Manafi, M., penyunting, InTechOpen, 2011, vol. 21, pp. 115–122.

  17. Papillon-Smith, J., Baker, S.E., Agbo, C., and Dahan, M.H., Pregnancy rates with intrauterine insemination: comparing 1999 and 2010 World Health Organization semen analysis norms, Reprod. Biomed. Online, 2015, vol. 30, pp. 392–400.

    Article  CAS  Google Scholar 

  18. Ricci, G., Perticarari, S., Boscolo, R., et al., Semen preparation methods and sperm apoptosis: swim-up versus gradient-density centrifugation technique, Fertil. Steril., 2009, vol. 91, pp. 632–638.

    Article  Google Scholar 

  19. Oseguera-López, I., Ruiz-Díaz, S., Ramos-Ibeas, P., et al., Novel techniques of sperm selection for improving IVF and ICSI outcomes, Front. Cell Dev. Biol., 2019, vol. 29, pp. 297–298.

    Google Scholar 

  20. Oguz, Y., Guler, I., Erdem, A., et al., The effect of swim-up and gradient sperm preparation techniques on deoxyribonucleic acid (DNA) fragmentation in subfertile patients, J. Assist. Reprod. Genet., 2018, vol. 35, pp. 1083–1089.

    Article  Google Scholar 

  21. Špaleková, E., Makarevich, A., and Pivko, J., Effect of caffeine on parameters of ram sperm motility, Slovak J. Anim. Sci., 2011, vol. 44, pp. 78–83.

    Google Scholar 

  22. Somfai, T., Bodó, S., Nagy, S., et al., Effect of swim up and Percoll treatment on viability and acrosome integrity of frozen-thawed bull spermatozoa, Reprod. Domest. Anim., 2002, vol. 37, pp. 285–290.

    Article  CAS  Google Scholar 

  23. Swain, J.E., Lai, D., Takayama, S., et al., Thinking big by thinking small: application of microfluidic technology to improve ART, Lab Chip, 2013, vol. 13, pp. 1213–1224.

    Article  CAS  Google Scholar 

  24. Talwar, P., Semen preparation in IVF, in Jaypee’s Video Atlas of Assisted Reproductive Technologies and Clinical Embryology, 2013, p. 189.

  25. Van der Ven, H.H., Jeyendran, R.S., Al-Hasani, S., et al., Glass wool column filtration of human semen: relation to swim-up procedure and outcome of IVF, Hum. Reprod., 1988, vol. 3, pp. 85–88.

    Article  CAS  Google Scholar 

  26. Vasudevan, K., Raber, J., and Sztein, J., Fertility comparison between wild type and transgenic mice by in vitro fertilization, Transgen. Res., 2010, vol. 19, pp. 587–594.

    Article  CAS  Google Scholar 

  27. Zhang, X.D., Chen, M.Y., Gao, Y., et al., The effects of different sperm preparation methods and incubation time on the sperm DNA fragmentation, Hum. Fertil., 2011, vol. 14, pp. 187–191.

    Article  CAS  Google Scholar 

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Funding

This study was financially supported by the School of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran (research project no. MUK.1394.220).

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Authors and Affiliations

  1. Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

    Omid Banafshi, Farzad Soleimani & Fardin Fathi

  2. Department of Medical Biology and Central Electron Microscope Laboratory, Medical Scool, Pecs University, Pecs, Hungary

    Hamid Ahmadi

  3. Department of Biological Sciences, School of Natural Sciences, University of Tabriz, Tabriz, Iran

    Farzad Soleimani

  4. Department of Basic Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

    Kaveh Rahimi

  5. Zoonoses Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

    Omid Banafshi & Kaveh Rahimi

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  1. Omid Banafshi
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  2. Hamid Ahmadi
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Correspondence to Fardin Fathi.

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Conflict of interest. The authors declare that they have no conflicts of interest.

Statement on the welfare of animals. This experimental animal study was conducted in terms of the guidelines of the Institutional Animal Care and Use Committee of Kurdistan University of Medical Sciences.

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Omid Banafshi, Ahmadi, H., Soleimani, F. et al. Sperm Preparation by Washing and Swim-up Method Improves In Vitro Fertilization and Birth Rate in Mice. Russ J Dev Biol 52, 449–456 (2021). https://doi.org/10.1134/S1062360421060102

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  • DOI: https://doi.org/10.1134/S1062360421060102

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