Emerging technologies in sperm selection: Can we solve ARTs frustration?
Sperm selection is a significant step in ARTs execution. We have already discussed the practiced methods for quality sperm selection. Several studies claim that fragmented sperm cells are able to fertilize female eggs but result in defective embryos. Our previous post advocates the importance of the sperm swimming mechanism. Except this, we highlighted here some promising techniques that have the potential to replace conventional sperm selection methods.
MAGNETIC ACTIVATED CELL SORTING (MACS)
MACS usually utilizes the magnetic field in separating apoptotic and no-apoptotic spermatozoa (Oseguera-López et al., 2019; Plouffe et al., 2015). Apoptosis is a process that comprises the biochemical transformation of the cells, which eventually end up in cell death. MACS comprises the labeling of sperm cells with superparamagnetic beads (~50 nm of Annexin V). However, labeling of Annexin V with sperm cell affect the membrane integrity (Agarwal et al., 2014).
The sperm sample mixed with Annexin V in a 1:1 ratio and incubated at room temperature for 15 minutes. The mixture is loaded into the MACS device where the 1.5 Tesla is applied across the sample holder. All Annexin V negative cells pass through the pellet and positive entangled in the columns. Annexin V negative cells are tested for motility and kept for cryopreservation (Agarwal et al., 2014).
HYALURONIC ACID (HA) BINDING
Female oocytes are surrounded by hyaluronic acid. Sperm cell comprises the receptors for hyaluronic binding often shows normal morphology and better DNA integrity (Oseguera-López et al., 2019). A Petri dish coated with hyaluronic acid is utilized to separate the responsive sperm cell. The entangled sperm cells at the bottom of the dish are taken further for the motility analysis (Nasr-Esfahani et al., 2008). However, the outcomes show equivalent integrity against the swim-up and centrifugation method (Mongkolchaipak and Vutyavanich, 2013).
ZETA METHOD
Usually, the membrane of mature sperm cells is negatively charged (Engelmann et al., 1988). The method comprises the two compartments separated with a porous membrane — one compartment with the semen sample and the other with sperm-free medium. An electric field is implemented across the terminal wall (anode at the sperm-free compartment). The negatively charged sperm membrane migrates towards the sperm-free compartment (Oseguera-López et al., 2019). Further motility analysis is revealed that collected sperm with the zeta method represent better DNA integrity against sperm swim-up, density centrifugation, and HA binding method (Razavi et al., 2010).
SELECTION BASED ON MORPHOLOGY
The normal morphology is one of the critical criteria in quality sperm selection. Since microscopy evolves, the manual selection and screening of sperm samples involve the percentage of normal morphology is one of the screening parameters. The development of high-resolution microscopy opens the opportunities for analyzing sperm morphology further.
Bartoove et al. 2002 introduced the technology “Motile Sperm Organelle Morphology Examination (MSOME)”, where high-resolution microscopy at 6300X involves selecting sperm cells with the low numbers vacuoles (Bartoov et al., 2002). Wilding et al. added that the sperm head involving large numbers of vacuoles corresponds to DNA fragmentation (Wilding et al., 2011). However, Leandri et al. not achieved a significant difference between the sperm selected with ICSI and MSOME (Leandri et al., 2013). Moreover, the technique is expensive and required a skilled workforce for execution.
BASED ON SWIMMING MECHANISM: MICROFLUIDICS
Currently, microfluidics-based technologies are being exploited to replicate the microenvironment and physiological conditions of the human organ. The method Researchers are replicating the natural mechanism utilizing the microfluidic toolboxes. Microfluidic-based technology facilitates high degree microenvironment control, including material transportation (mass flow, diffusion, and mixing).
The mechanism of sperm swimming involves rheotaxis (sperm swimming upstreams (El-Sherry et al., 2014; Elsayed et al., 2015; Kantsler et al., 2014; Zaferani et al., 2018)), thermotaxis (sperm migration towards through temperature gradient (Bahat et al., 2012, 2003; Bahat and Eisenbach, 2006)), and chemotaxis (sperm migration towards through chemical gradient (Bhagwat et al., 2018; Eisenbach, 1999; Koyama et al., 2006; Xie et al., 2010)). However, the translation rate of these technologies is slow; moreover, the user-friendliness, material choice, and high throughput appeared as significant constraints in commercialization pathways.
REFERENCES
Agarwal, A., Sharma, R., Beydola, T., 2014. Sperm Preparation and Selection Techniques, in: Medical and Surgical Management of Male Infertility. https://doi.org/10.5005/jp/books/11840_29
Bahat, A., Caplan, S.R., Eisenbach, M., 2012. Thermotaxis of human sperm cells in extraordinarily shallow temperature gradients over a wide range. PLoS One. https://doi.org/10.1371/journal.pone.0041915
Bahat, A., Eisenbach, M., 2006. Sperm thermotaxis. Mol. Cell. Endocrinol. https://doi.org/10.1016/j.mce.2006.03.027
Bahat, A., Tur-Kaspa, I., Gakamsky, A., Giojalas, L.C., Breitbart, H., Eisenbach, M., 2003. Thermotaxis of mammalian sperm cells: A potential navigation mechanism in the female genital tract [1]. Nat. Med. https://doi.org/10.1038/nm0203-149
Bartoov, B., Berkovitz, A., Eltes, F., Kogosowski, A., Menezo, Y., Barak, Y., 2002. Real-time fine morphology of motile human sperm cells is associated with IVF-ICSI outcome. J. Androl. 23. https://doi.org/10.1002/j.1939-4640.2002.tb02595.x
Bhagwat, S., Sontakke, S., Deekshith, K., Parte, P., Jadhav, S., 2018. Chemotactic behavior of spermatozoa captured using a microfluidic chip. Biomicrofluidics. https://doi.org/10.1063/1.5023574
Eisenbach, M., 1999. Sperm chemotaxis. Rev. Reprod. https://doi.org/10.1530/ror.0.0040056
El-Sherry, T.M., Elsayed, M., Abdelhafez, H.K., Abdelgawad, M., 2014. Characterization of rheotaxis of bull sperm using microfluidics. Integr. Biol. (United Kingdom). https://doi.org/10.1039/c4ib00196f
Elsayed, M., El-Sherry, T.M., Abdelgawad, M., 2015. Development of computer-assisted sperm analysis plugin for analyzing sperm motion in microfluidic environments using Image-J. Theriogenology. https://doi.org/10.1016/j.theriogenology.2015.07.021
Engelmann, U., Krassnigg, F., Schatz, H., Schill, W. ‐B, 1988. Separation of human X and Y spermatozoa by free‐flow electrophoresis. Gamete Res. 19. https://doi.org/10.1002/mrd.1120190205
Kantsler, V., Dunkel, J., Blayney, M., Goldstein, R.E., 2014. Rheotaxis facilitates upstream navigation of mammalian sperm cells. Elife. https://doi.org/10.7554/elife.02403
Koyama, S., Amarie, D., Soini, H.A., Novotny, M. V., Jacobson, S.C., 2006. Chemotaxis assays of mouse sperm on microfluidic devices. Anal. Chem. https://doi.org/10.1021/ac052087i
Leandri, R.D., Gachet, A., Pfeffer, J., Celebi, C., Rives, N., Carre-Pigeon, F., Kulski, O., Mitchell, V., Parinaud, J., 2013. Is intracytoplasmic morphologically selected sperm injection (IMSI) beneficial in the first ART cycle? A multicentric randomized controlled trial. Andrology 1. https://doi.org/10.1111/j.2047-2927.2013.00104.x
Mongkolchaipak, S., Vutyavanich, T., 2013. No difference in high-magnification morphology and hyaluronic acid binding in the selection of euploid spermatozoa with intact DNA. Asian J. Androl. 15. https://doi.org/10.1038/aja.2012.163
Nasr-Esfahani, M.H., Razavi, S., Vahdati, A.A., Fathi, F., Tavalaee, M., 2008. Evaluation of sperm selection procedure based on hyaluronic acid binding ability on ICSI outcome. J. Assist. Reprod. Genet. 25. https://doi.org/10.1007/s10815-008-9223-4
Oseguera-López, I., Ruiz-Díaz, S., Ramos-Ibeas, P., Pérez-Cerezales, S., 2019. Novel Techniques of Sperm Selection for Improving IVF and ICSI Outcomes. Front. Cell Dev. Biol. https://doi.org/10.3389/fcell.2019.00298
Plouffe, B.D., Murthy, S.K., Lewis, L.H., 2015. Fundamentals and application of magnetic particles in cell isolation and enrichment: A review. Reports Prog. Phys. https://doi.org/10.1088/0034-4885/78/1/016601
Razavi, S.H., Nasr-Esfahani, M.H., Deemeh, M.R., Shayesteh, M., Tavalaee, M., 2010. Evaluation of zeta and HA-binding methods for selection of spermatozoa with normal morphology, protamine content and DNA integrity. Andrologia 42. https://doi.org/10.1111/j.1439-0272.2009.00948.x
Wilding, M., Coppola, G., Di Matteo, L., Palagiano, A., Fusco, E., Dale, B., 2011. Intracytoplasmic injection of morphologically selected spermatozoa (IMSI) improves outcome after assisted reproduction by deselecting physiologically poor quality spermatozoa. J. Assist. Reprod. Genet. 28. https://doi.org/10.1007/s10815-010-9505-5
Xie, L., Ma, R., Han, C., Su, K., Zhang, Q., Qiu, T., Wang, L., Huang, G., Qiao, J., Wang, J., Cheng, J., 2010. Integration of sperm motility and chemotaxis screening with a microchannel-based device. Clin. Chem. https://doi.org/10.1373/clinchem.2010.146902
Zaferani, M., Cheong, S.H., Abbaspourrad, A., 2018. Rheotaxis-based separation of sperm with progressive motility using a microfluidic corral system. Proc. Natl. Acad. Sci. U. S. A. https://doi.org/10.1073/pnas.1800819115
