Sperm Cryopreservation

  • Takeshi ShinEmail author
  • Mai Fukushima
  • Akane Miyata
  • Hiroshi Okada


Sperm cryopreservation, the preservation and storage of sperm for future use, is an efficient procedure for the management of male fertility and is now routinely offered to men as a means of preserving their fertility. Cryopreservation should be discussed and offered prior to cytotoxic therapy for not only malignant diseases but also nonmalignant systemic diseases such as autoimmune disorders or inflammatory bowel disease. There are many important steps in the process of successful cryopreservation. Freezing and thawing should be performed using techniques that minimize sperm damage. Although sperm banking is now a common technique all over the world, little has been written about the clinical process. In this chapter, we provide an overview of sperm cryopreservation, including its history and the methods used for cryopreservation. We also provide the indications of sperm cryopreservation and scientific background of the procedures.


Cryopreservation Fertility preservation History Sperm 


  1. 1.
    Anger JT, Gilbert BR, Goldstein M (2003) Cryopreservation of sperm: indications, methods and results. J Urol 170:1079–1084CrossRefPubMedGoogle Scholar
  2. 2.
    Bunge RG, Keettel WC, Sherman JK (1954) Clinical use of frozen semen: report of four cases. Fertil Steril 5:520–529CrossRefPubMedGoogle Scholar
  3. 3.
    Walters EM, Benson JD, Woods EJ et al (2009) The history of sperm cryopreservation. In: Pacey AA, Tomlinson M (eds) Sperm banking theory and practice. Cambridge University Press, London, pp 1–17Google Scholar
  4. 4.
    Polge C, Smith AU, Parkes AS (1949) Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature 164:666CrossRefPubMedGoogle Scholar
  5. 5.
    Polge C (1952) Fertilizing capacity of bull spermatozoa after freezing at −79°C. Nature 169:626–627CrossRefPubMedGoogle Scholar
  6. 6.
    Bunge RG, Sherman JK (1953) Fertilizing capacity of frozen human spermatozoa. Nature 172:767–768CrossRefPubMedGoogle Scholar
  7. 7.
    Sherman JK (1980) Historical synopsis of human semen cryobanking. In: David G, Price WS (eds) Human artificial insemination and semen preservation. Springer, New York, pp 95–105CrossRefGoogle Scholar
  8. 8.
    Hamada A, Wasik M, Gupta S et al (2012) Sperm banking: indications and regulations. In: Dubey AK (ed) Infertility: diagnosis, management and IVF. Jaypee Brothers Medical Publishing, New Delhi, pp 409–434CrossRefGoogle Scholar
  9. 9.
    Sabanegh ES Jr, Ragheb AM (2009) Male fertility after cancer. Urology 73:225–231CrossRefPubMedGoogle Scholar
  10. 10.
    Tomlinson M (2010) Therapeutic sperm cryopreservation. In: Björndahl L, Giwercman A, Tournaye L (eds) Clinical andrology EAU/ESAU course guidelines. Informa Health Care, London, pp 124–133CrossRefGoogle Scholar
  11. 11.
    Meistrich ML, Finch M, da Cunha MF et al (1982) Damaging effects of fourteen chemotherapeutic drugs on mouse testis cells. Cancer Res 42:122–131PubMedGoogle Scholar
  12. 12.
    Sigman M (2013) Cancer treatment and male infertility: effects of therapy and current and future management options. Fertil Steril 100:1179–1186CrossRefPubMedGoogle Scholar
  13. 13.
    Meistrich ML, Wilson G, Mathur K et al (1997) Rapid recovery of spermatogenesis after mitoxantrone, vincristine, vinblastine, and prednisone chemotherapy for Hodgkin’s disease. J Clin Oncol 15:3488–3495PubMedGoogle Scholar
  14. 14.
    Meistrich ML (2009) Effect of antineoplastic and other medical treatments on sperm production. In: Pacey AA, Tomlinson M (eds) Sperm banking theory and practice. Cambridge University Press, London, pp 18–29Google Scholar
  15. 15.
    Gordon W, Siegmund K, Stanisic TH et al (1997) A study of reproductive function in patients with seminoma treated with radiotherapy and orchidectomy: (SWOG-8711). Int J Radiat Oncol Biol Phys 38:83–94CrossRefPubMedGoogle Scholar
  16. 16.
    Wallace WH (2011) Oncofertility and preservation of reproductive capacity in children and young adults. Cancer 117:2301–2310CrossRefPubMedGoogle Scholar
  17. 17.
    Colpi GM, Contalbi GF, Nerva F et al (2004) Testicular function following chemo-radiotherapy. Eur J Obstet Gynecol Reprod Biol 113:S2–S6CrossRefPubMedGoogle Scholar
  18. 18.
    Ranganathan P, Mahran AM, Hallak J et al (2002) Sperm cryopreservation for men with nonmalignant, systemic diseases: a descriptive study. J Androl 23:71–75PubMedGoogle Scholar
  19. 19.
    Mishra DP, Shaha C (2005) Estrogen-induced spermatogenic cell apoptosis occurs via the mitochondrial pathway: role of superoxide and nitric oxide. J Biol Chem 280:6181–6196CrossRefPubMedGoogle Scholar
  20. 20.
    Gupta S, Agarwal A, Sharma R (2011) Recovery, preparation, storage and utilization of spermatozoa for fertility preservation in cancer patients and subfertile men. J Reprod Stem Cell Biotechnol 1:150–168Google Scholar
  21. 21.
    World Health Organization, Department of Reproductive Health and Research (2010) WHO laboratory manual for the examination and processing of human semen, 5th edn.Google Scholar
  22. 22.
    Okada H, Dobashi M, Yamazaki T et al (2002) Conventional versus microdissection testicular sperm extraction for nonobstructive azoospermia. J Urol 168:1063–1067CrossRefPubMedGoogle Scholar
  23. 23.
    Marmar JL (1998) The emergence of specialized procedures for the acquisition, processing, and cryopreservation of epididymal and testicular sperm in connection with intracytoplasmic sperm injection. J Androl 19:517–526PubMedGoogle Scholar
  24. 24.
    Kupker W, Schlegel PN, Al-Hasani S et al (2000) Use of frozen-thawed testicular sperm for intracytoplasmic sperm injection. Fertil Steril 73:453–458CrossRefPubMedGoogle Scholar
  25. 25.
    Bak CW, Song SH, Yoon TK et al (2010) Natural course of idiopathic oligozoospermia: comparison of mild, moderate and severe forms. Int J Urol 17:937–943CrossRefPubMedGoogle Scholar
  26. 26.
    Song SH, Bak CW, Lim JJ et al (2010) Natural course of severe oligozoospermia in infertile male: influence on future fertility potential. J Androl 31:536–539CrossRefPubMedGoogle Scholar
  27. 27.
    Practice Committee of American Society for Reproductive Medicine (2013) Recommendations for reducing the risk of viral transmission during fertility treatment with the use of autologous gametes: a committee opinion. Fertil Steril 99:340–346CrossRefGoogle Scholar
  28. 28.
    Tomlinson MJ (2009) Sperm processing and storage. In: Pacey AA, Tomlinson M (eds) Sperm banking theory and practice. Cambridge University Press, London, pp 86–104Google Scholar
  29. 29.
    Kalthur G, Adiga SK, Upadhya D et al (2008) Effect of cryopreservation on sperm DNA integrity in patients with teratospermia. Fertil Steril 89:1723–1727CrossRefPubMedGoogle Scholar
  30. 30.
    Sherma RK, Agarwal A (1997) Influence of artificial stimulation on unprocessed and Percoll-washed cryopreserved sperm. Arch Androl 38:173–179CrossRefGoogle Scholar
  31. 31.
    Esteves SC, Sharma RK, Thomas AJ Jr et al (2000) Improvement in motion characteristics and acrosome status in cryopreserved human spermatozoa by swim-up processing before freezing. Human Reprod 15:2173–2179CrossRefGoogle Scholar
  32. 32.
    Behrman SJ, Sawada Y (1966) Heterologous and homologous inseminations with human semen frozen and stored in a liquid-nitrogen refrigerator. Fertil Steril 17:457–466CrossRefPubMedGoogle Scholar
  33. 33.
    Di Santo M, Tarozzi N, Nadalini M et al (2012) Human sperm cryopreservation: update on techniques, effect on DNA integrity, and implications for ART. Adv Urol 2012:854837CrossRefPubMedGoogle Scholar
  34. 34.
    Mahadevan M, Trounson AO (1984) Effect of cooling, freezing, and thawing rates and storage conditions on preservation of human spermatozoa. Andrologia 16:52–60CrossRefPubMedGoogle Scholar
  35. 35.
    Mazur P, Rall WF, Rigopoulos N (1981) Relative contributions of the fraction of unfrozen water and of salt concentration to the survival of slowly frozen human erythrocytes. Biophys J 36:653–675CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Nallella KP, Sharma RK, Allamaneni SS et al (2004) Cryopreservation of human spermatozoa: comparison of two cryopreservation methods and three cryoprotectants. Fertil Steril 82:913–918CrossRefPubMedGoogle Scholar
  37. 37.
    Fahy GM, MacFarlane DR, Angell CA et al (1984) Vitrification as an approach to cryopreservation. Cryobiology 21:407–426CrossRefPubMedGoogle Scholar
  38. 38.
    Cohen J, Garrisi GJ, Congedo-Ferrara TA et al (1997) Cryopreservation of single human spermatozoa. Hum Reprod 12:994–1001CrossRefPubMedGoogle Scholar
  39. 39.
    Critser JK, Huse-Benda AR, Aaker DV et al (1988) Cryopreservation of human spermatozoa. III. The effect of cryoprotectants on motility. Fertil Steril 50:314–320CrossRefPubMedGoogle Scholar
  40. 40.
    Hammitte DG, Walker DL, Williamson RA (1988) Concentration of glycerol required for optimal survival and in vitro fertilizing capacity of frozen sperm is dependent on cryopreservation medium. Fertil Steril 4:680–687CrossRefGoogle Scholar
  41. 41.
    Jeyendran RS, Van der Ven MM, Kennedy W et al (1984) Comparison of glycerol and a zwitterion buffer system as cryoprotective media for human spermatozoa. J Androl 5:1–6CrossRefPubMedGoogle Scholar
  42. 42.
    Jeyendran RS, Van der Ven MM, Kennedy W et al (1985) Effect of glycerol and cryopreservation on oocyte penetration by human spermatozoa. Andrologia 17:241–248CrossRefPubMedGoogle Scholar
  43. 43.
    Puris GS, Weidel L (1986) A comparative study of buffer systems as cryoprotectants for human spermatozoa. Fertil Steril 46:147–149CrossRefGoogle Scholar
  44. 44.
    Mortimer D (2004) Current and future concepts and practices in human sperm cryobanking. Reprod Biomed Online 9:134–151CrossRefPubMedGoogle Scholar

Copyright information

© Springer Japan 2016

Authors and Affiliations

  • Takeshi Shin
    • 1
    Email author
  • Mai Fukushima
    • 1
  • Akane Miyata
    • 1
  • Hiroshi Okada
    • 1
  1. 1.Center for Reproductive MedicineDokkyo Medical University Koshigaya HospitalKoshigayaJapan

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