Skip to main content Accessibility help

We use cookies to distinguish you from other users and to provide you with a better experience on our websites. Close this message to accept cookies or find out how to manage your cookie settings.

Close cookie message
×
Hostname: page-component-cd9895bd7-dk4vv Total loading time: 0 Render date: 2024-12-26T15:11:29.576Z Has data issue: false hasContentIssue false

Chapter 12 - Sperm Cryopreservation

from Part III - Laboratory Handling of Retrieved Sperm

Published online by Cambridge University Press:  09 April 2021

By
Rakesh Sharma ,
Kruyanshi Master  and
Ashok Agarwal
Edited by
Ashok Agarwal ,
Ahmad Majzoub  and
Sandro C. Esteves
Ashok Agarwal
Affiliation:
The Cleveland Clinic Foundation, Cleveland, OH, USA
Ahmad Majzoub
Affiliation:
Hamad Medical Corporation, Doha, Qatar
Sandro C. Esteves
Affiliation:
Andrology & Human Reproduction Clinic, Sao Paulo, Brazil
Get access

Summary

Human sperm cryopreservation is a highly desirable technique for preserving fertility potential and future use in couples desiring to have a biological child. Slow cryopreservation of sperm has been the mainstay technique. The drawback of this technique is the inability to freeze extremely small numbers of sperm as in the case of surgical retrieval of testicular sperm. These sperm are extremely few in number and it is difficult to retrieve motile, viable sperm post-thaw. In the past decade, sperm vitrification has been introduced along with both biological and non-biological carriers to freeze extremely low numbers of sperm. Vitrification also allows the ability to freeze single spermatozoa. These techniques are in many ways more efficient and better than the older techniques. This chapter aims to provide a detailed introduction to various approaches used for preserving spermatozoa. It also discusses indications of sperm cryopreservation based on semen quality and summarizes the advantages and shortcomings of these techniques.

Keywords

spermsperm cryopreservationslow freezingvitrificationsperm storagesperm motilityDNA integrity
Type
Chapter
Information
Manual of Sperm Retrieval and Preparation in Human Assisted Reproduction , pp. 99 - 116
Publisher: Cambridge University Press
Print publication year: 2021

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

AbdelHafez, F, Bedaiwy, M, El-Nashar, SA, Sabanegh, E, Desai, N. Techniques for cryopreservation of individual or small numbers of human spermatozoa: a systematic review. Hum Reprod Update 2008;15:153–164.Google Scholar
Holt, W. Basic aspects of frozen storage of semen. Anim Reprod Sci 2000;62(1–3):3–22.Google Scholar
Hezavehei, M, Sharafi, M, Kouchesfahani, HM, et al. Sperm cryopreservation: a review on current molecular cryobiology and advanced approaches. Reprod Biomed Online 2018;37(3):327–339.CrossRefGoogle ScholarPubMed
Agha‐Rahimi, A, Khalili, M, Nottola, S, Miglietta, S, Moradi, A. Cryoprotectant‐free vitrification of human spermatozoa in new artificial seminal fluid. Andrology 2016;4(6):1037–1044.CrossRefGoogle ScholarPubMed
Di Santo, M, Tarozzi, N, Nadalini, M, Borini, A. Human sperm cryopreservation: update on techniques, effect on DNA integrity, and implications for ART. Adv Urol 2012;2012:854837.Google Scholar
Mocé, E, Fajardo, AJ, Graham, JK. Human sperm cryopreservation. EMJ 2016;1(1):86–91.CrossRefGoogle Scholar
Hammerstedt, RH, Graham, JK, Nolan, JP. Cryopreservation of mammalian sperm: what we ask them to survive. J Androl 1990;11(1):73–88.Google Scholar
Mazur, P. Freezing of living cells: mechanisms and implications. Am J Physiol Cell Physiol 1984;247(3):C125–C142.CrossRefGoogle ScholarPubMed
Day, JG, Harding, KC, Nadarajan, J, Benson, EE. Cryopreservation: Molecular Biomethods Handbook. Humuna Press, Totowa, NJ, 2008.Google Scholar
Rozati, H, Handley, T, Jayasena, C. Process and pitfalls of sperm cryopreservation. J Clin Med 2017;6(9):89.Google Scholar
Medrano, JV, Del Mar Andrés, M, García, S, et al. Basic and clinical approaches for fertility preservation and restoration in cancer patients. Trends Biotechnol 2018;36(2):199–215.CrossRefGoogle ScholarPubMed
Gupta, S, Agarwal, A, Sharma, R, Ahmady, A. Sperm banking via cryopreservation. In: RizkBotros, RMB, Aziz, N, Agarwal, A, Sabanegh, E (eds.) Medical and Surgical Management of Male Infertility. Jaypee Brothers, New Delhi, 2014, pp. 234–243.Google Scholar
Kagalwala, S. Sperm vitrification. In Allahabadia, G, Kuwayama, M, Gandhi, G (eds.) Vitrification in Assisted Reproduction: A User’s Manual. Springer, New Delhi, 2015, pp. 31–42.Google Scholar
Agarwal, A, Gupta, S, Sharma, R. Cryopreservation of client depositor semen. In: Agarwal, A, Gupta, S, Sharma, R (eds.) Andrological Evaluation of Male Infertility: A Laboratory Guide. Springer, Cham, 2016, pp. 113–133.Google Scholar
Agarwal, A, Tvrda, E. Slow freezing of human sperm. In: Nagy Zsolt, P, Vargheses, A, Agarwal, A (eds.) Cryopreservation of Mammalian Gametes and Embryos: Methods and Protocols. Springer Science + Business Media, New York, 2017, pp. 67–78.Google Scholar
Sharma, R, Kattoor, AJ, Ghulmiyyah, J, Agarwal, A. Effect of sperm storage and selection techniques on sperm parameters. Syst Biol Reprod Med 2015;61(1):1–12.Google Scholar
Gupta, S, Sharma, R, Agarwal, A. The process of sperm cryopreservation, thawing and washing techniques. In: Majoub, A, Agarwal, A (eds.) The Complete Guide to Male Fertility Preservation. Springer, Cham, 2018, pp. 183–204.Google Scholar
Agarwal, A, Sharma, R, Gupta, S, Sharma, R. NextGen® home sperm banking kit: outcomes of offsite vs onsite collection—preliminary findings. Urology 2015;85(6):1339–1346.Google Scholar
Bunge, R, Sherman, J. Fertilizing capacity of frozen human spermatozoa. Nature 1953;172(4382):767–768.CrossRefGoogle ScholarPubMed
Oldenhof, HM. Gojowsky, S, Wang, S, et al. Osmotic stress and membrane phase changes during freezing of stallion sperm: mode of action of cryoprotective agents. Biol Reprod 2013;88(3):68.Google Scholar
Isachenko, V, Maettner, R, Petrunkina, A, et al. Cryoprotectant-free vitrification of human spermatozoa in large (up to 0.5 mL) volume: a novel technology. Clin Lab 2011;57(9–10):643–650.Google Scholar
Mohamed, MSA. Slow cryopreservation is not superior to vitrification in human spermatozoa; an experimental controlled study. Iranian J Reprod Med 2015;13(10):633–644.Google Scholar
Le, MT, Nguyen, TTT, Nguyen, TT, et al. Cryopreservation of human spermatozoa by vitrification versus conventional rapid freezing: effects on motility, viability, morphology and cellular defects. Eur J Obstet Gynecol Reprod Biol 2019;234:14–20.CrossRefGoogle ScholarPubMed
Li, YX, Zhou, L, LV MQ, et al. Vitrification and conventional freezing methods in sperm cryopreservation: a systematic review and meta-analysis. Eur J Obstet Gynecol Reprod Biol 2019;233:84–92.CrossRefGoogle ScholarPubMed
Slabbert, M, Du Plessis, S, Huyser, C. Large volume cryoprotectant‐free vitrification: an alternative to conventional cryopreservation for human spermatozoa. Andrologia 2015;47(5):594–599.Google Scholar
Aizpurua, J, Medrano, L, Enciso, M, et al. New permeable cryoprotectant-free vitrification method for native human sperm. Hum Reprod 2017;32(10):2007–2015.Google Scholar
Isachenko, E, Isachenko, V, Katkov, II, Dessole, S, Nawroth, F. Vitrification of mammalian spermatozoa in the absence of cryoprotectants: from past practical difficulties to present success. Reprod Biomed Online 2003;6(2):191–200.CrossRefGoogle ScholarPubMed
Isachenko, E, Isachenko, V, Katkov, II et al. DNA integrity and motility of human spermatozoa after standard slow freezing versus cryoprotectants-free vitrification. Hum Reprod 2004;19:932–939.Google Scholar
Isachenko, V, Isachenko, E, Katkov, II et al. Cryoprotectant-free cryopreservation of human spermatozoa by vitrification and freezing in vapor: effect of motility, DNA integrity, and fertilization ability. Biol Reprod 2004;71:1167–1173.Google Scholar
Mansilla, MA, Merino, O, Risopatron, J, et al. High temperature is essential for preserved human sperm function during the devitrification process. Andrologia 2016;48:111–113.Google Scholar
Khalili, MA, Adib, M, Halvaei, I, Nabi, A. Vitrification of neat semen alters sperm parameters and DNA integrity. Urol J 2014;11(2):1465–1470.Google Scholar
Riel, JM, Yamauchi, Y, Huang, TT, Grove, J, Ward, MA. Short-term storage of human spermatozoa in electrolyte-free medium without freezing maintains sperm chromatin integrity better than cryopreservation. Biol Reprod 2011;85(3):536–547.Google Scholar
Podsiadly, BT, Woolcott, RJ, Stanger, JD, Stevenson, K. Pregnancy resulting from intracytoplasmic injection of cryopreserved spermatozoa recovered from testicular biopsy. Hum Reprod 1996;11(6):1306–1308.Google Scholar
Cohen, J, Garrisi, GJ, Congedo-Ferrara, TA, et al. Cryopreservation of single human spermatozoa. Hum Reprod 1997;12(5):994–1001.Google Scholar
Isachenko, V. Clean technique for cryoprotectant-free vitrification of human spermatozoa. RBM Online 2005;10(3):350–354.Google Scholar
Gil-Salom, MJ, Romero, C, Rubio, A, Remohı, RJ, Pellicer, A. Intracytoplasmic sperm injection with cryopreserved testicular spermatozoa. Mol Cell Endocrinol 2000;169(1-2):15–19.Google Scholar
Lane, M, Bavister, BD, Lyons, EA, Forest, KT. Containerless vitrification of mammalian oocytes and embryos. Nat Biotechnol 1999;17(12):1234–1236.Google Scholar
Schuster, TG, Keller, LM, Dunn, RL, Ohl, DA, Smith, GD. Ultra‐rapid freezing of very low numbers of sperm using cryoloops. Hum Reprod 2003;18(4):788–795.Google Scholar
Endo, Y, Fujii, Y, Shintani, K, et al. Simple vitrification for small numbers of human spermatozoa. Reprod Biomed Online 2012;24:301–307.Google Scholar
Royere, D, Hamamah, S, Nicolle, J, Lansac, J. Chromatin alterations induced by freeze–thawing influence the fertilizing ability of human sperm. Int J Androl 1991;14(5):328–332.Google Scholar
Gangrade, BK. Cryopreservation of testicular and epididymal sperm: techniques and clinical outcomes of assisted conception. Clinics 2013;68:131–140.CrossRefGoogle ScholarPubMed
Friedler, S, Strassburger, D, Raziel, A, et al. Intracytoplasmic injection of fresh and cryopreserved testicular spermatozoa in patients with nonobstructive azoospermia: a comparative study. Fertil Steril 1997;68(5):892–897.Google Scholar
Moskovtsev, S, Lulat, A, Librach, C. Cryopreservation of Human Spermatozoa by Vitrification vs. Slow Freezing: Canadian Experience. Intech Open, London, 2012.Google Scholar
Schlegel, PN. Testicular sperm extraction: microdissection improves sperm yield with minimal tissue excision. Hum Reprod 1999;14:131–135.Google Scholar
Esteves, SC; Miyaoka, R, Agarwal, A. Sperm retrieval techniques for assisted reproduction. Int Braz J Urol 2011; 37(5): 570–583.Google Scholar
Desai, N, Blackmon, H, Goldfarb, J. Single sperm cryopreservation on cryoloops: an alternative to hamster zona for cryopreservation of individual spermatozoa. Fertil Steril 2003;80:55–56.Google Scholar
Tournaye, H, Merdad, T, Silber, S, et al. No differences in outcome after intracytoplasmic sperm injection with fresh or with frozen–thawed epididymal spermatozoa. Hum Reprod 1999;14(1):90–95.Google Scholar
Cayan, S, Lee, D, Conaghan, J, et al. A comparison of ICSI outcomes with fresh and cryopreserved epididymal spermatozoa from the same couples. Hum Reprod 2001;16(3):495–499.Google Scholar
Shibahara, H, Hamada, Y, Hasegawa, A, et al. Correlation between the motility of frozen-thawed epididymal spermatozoa and the outcome of intracytoplasmic sperm injection. Int J Androl 1999;22(5):324–328.Google Scholar
Kuczynski, W, Dhont, M, Grygoruk, C, et al. The outcome of intracytoplasmic injection of fresh and cryopreserved ejaculated spermatozoa: a prospective randomized study. Hum Reprod 2001;16(10):2109–2113.Google Scholar
Ragni, G, Caccamo, AM, Dalla Serra, A, Guercilena, S. Computerized slow-staged freezing of semen from men with testicular tumors or Hodgkin’s disease preserves sperm better than standard vapor freezing. Fertil Steril 1990;53(6):1072–1075.Google Scholar
Borges, E Jr, Rossi, LM, Locambo de Freitas, CV, et al. Fertilization and pregnancy outcome after intracytoplasmic injection with fresh or cryopreserved ejaculated spermatozoa. Fertil Steril 2007;87(2):316–320.Google Scholar
Gupta, S, Sekhon, LH, Agarwal, A. Sperm banking: when, why, and how? In: Sabanegh, ES Jr. (ed.) Male Infertility. Current Clinical Urology: Male Infertility: Problems and Solutions. Springer, Cham, 2011, pp. 107–118.Google Scholar
Kliesch, S, Kamischke, A, Cooper, TG, Nieschlag, E. Cryopreservation of human spermatozoa. In: Andrology, Springer, Cham, 2010, pp. 505–520.Google Scholar
Spanò, M, Cordelli, E, Leter, G, et al. Nuclear chromatin variations in human spermatozoa undergoing swim-up and cryopreservation evaluated by the flow cytometric sperm chromatin structure assay. Mol Hum Reprod 1999;5(1):29–37.CrossRefGoogle ScholarPubMed
Hammadeh, ME, Dehn, C, Hippach Zeginiadou, M, et al. Comparison between computerized slow-stage and static liquid nitrogen vapour freezing methods with respect to the deleterious effect on chromatin and morphology of spermatozoa from fertile and subfertile men. Int J Androl 2001;24(2):66–72.Google Scholar
Gandini, l, Lombardo, F, Lenzi, A, Spanò, M, Dondero, F. Cryopreservation and sperm DNA integrity. Cell Tissue Banking 2006;7(2):91–98.Google Scholar
Petyim, S, Choavaratana, R. Cryodamage on sperm chromatin according to different freezing methods, assessed by AO test. J Med Assoc Thailand 2006;89(3):306–313.Google Scholar
Ngamwuttiwong, T, Kunathikom, S. Evaluation of cryoinjury of sperm chromatin according to liquid nitrogen vapour method (I). J Med Assoc Thailand 2007;90(2):224–228.Google Scholar
Ahmad, L, Jalali, S, Shami, SA, et al. Effects of cryopreservation on sperm DNA integrity in normospermic and four categories of infertile males. Syst Biol Reprod Med 2010;56(1):74–83.Google Scholar
Donnelly, ET, McClure, N, Lewis, SE. Cryopreservation of human semen and prepared sperm: effects on motility parameters and DNA integrity. Fertil Steril 2001;76(5):892–900.Google Scholar
Kalthur, G, Adiga, SK, Upadhya, D, Rao, S, Kumar, P. Effect of cryopreservation on sperm DNA integrity in patients with teratospermia. Fertil Steril 2008;89(6):1723–1727.Google Scholar
de Paula, TS, Bertolla, RP, Spaine, DM, et al. Effect of cryopreservation on sperm apoptotic deoxyribonucleic acid fragmentation in patients with oligozoospermia. Fertil Steril 2006;86(3):597–600.Google Scholar
Thomson, LK, Fleming, SD, Aitken, RJ, et al. Cryopreservation-induced human sperm DNA damage is predominantly mediated by oxidative stress rather than apoptosis. Hum Reprod 2009;24(9):2061–2070.Google Scholar
Zribi, N, Chakroun, NF, El Euch, H, et al. Effects of cryopreservation on human sperm deoxyribonucleic acid integrity. Fertil Steril 2010;93(1):159–166.Google Scholar
Høst, E, Lindenberg, S, Kahn, JA, Christensen, F. DNA strand breaks in human sperm cells: a comparison between men with normal and oligozoospermic sperm samples. Acta Obstet Gynecol Scand 1999;78(4):336–339.CrossRefGoogle Scholar
Steele, EK, McClure, N, Lewis, SEM. Comparison of the effects of two methods of cryopreservation on testicular sperm DNA. Fertil Steril 2000;74(3):450–453.Google Scholar
Duru, NK, Morshedi, MS, Schuffner, A, Oehninger, S. Cryopreservation-thawing of fractionated human spermatozoa is associated with membrane phosphatidylserine externalization and not DNA fragmentation. J Androl 2001;22(4):646–651.Google Scholar
Paasch, U, Sharma, RK, Gupta, AK, et al. Cryopreservation and thawing is associated with varying extent of activation of apoptotic machinery in subsets of ejaculated human spermatozoa. Biol Reprod 2004;71(6):1828–1837.Google Scholar
Varghese, AC, Nandi, P, Mahfouz, R, Athayde, KS, Agarwal, A. Human Sperm Cryopreservation: Andrology Laboratory Manual. Jaypee Brothers, New Delhi, 2010, pp. 196–208.Google Scholar

Save book to Kindle

To save this book to your Kindle, first ensure no-reply@cambridge.org is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about saving to your Kindle.

Note you can select to save to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be saved to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

Find out more about the Kindle Personal Document Service.

Available formats
×

Save book to Dropbox

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Dropbox.

Available formats
×

Save book to Google Drive

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Google Drive.

Available formats
×