Sperm Cryopreservation Methods

  • Tiffany Justice
  • Greg ChristensenEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 927)


There are multiple clinical situations where cryopreservation of sperm is required including sperm banking prior to chemotherapy or radiation treatment for cancer, donor sperm for couples without a male partner, and various etiologies of male factor infertility. Bunge and Sherman presented the first pregnancy resulting from the thawing of cryopreserved sperm in 1953 (Bunge RJ, Nature 172:767–768, 1953). Since that time, cryopreservation techniques have continued to improve. Here, we describe protocols for the cryopreservation and thawing of semen and testicular tissues using a simple liquid nitrogen vapor technique.

Key words

Sperm Cryopreservation Sperm-banking Liquid nitrogen 


  1. 1.
    Bunge RJ (1953) Fertilizing capacity of frozen human spermatozoa. Nature 172:767–768PubMedCrossRefGoogle Scholar
  2. 2.
    Anger JT et al (2003) Cryopreservation of sperm: indications, methods and results. J Urol 170:1079–1084PubMedCrossRefGoogle Scholar
  3. 3.
    Thonneau P et al (1991) Incidence and main causes of infertility in a resident population (1,850,000) of three French regions (1988–1989). Hum Reprod 6:811–816PubMedGoogle Scholar
  4. 4.
    Khorram O et al (2001) Reproductive technologies for male infertility. J Clin Endocrinol Metab 86:2373–2379PubMedCrossRefGoogle Scholar
  5. 5.
    Isachenko V et al (2012) Vitrification of human ICSI/IVF spermatozoa without cryoprotectants: new capillary technology. J Androl 33(3):462–468Google Scholar
  6. 6.
    Isachenko V et al (2004) Cryoprotectant-free cryopreservation of human spermatozoa by vitrification and freezing in vapor: effect on motility. DNA integrity, and fertilization ability. Biol Reprod 71:1167–1173PubMedCrossRefGoogle Scholar
  7. 7.
    Rofeim O et al (2001) Effects of serial thaw-refreeze cycles on human sperm motility and viability. Fertil Steril 75:1242–1243PubMedCrossRefGoogle Scholar
  8. 8.
    Bandularatne E, Bongso A (2002) Evaluation of human sperm function after repeated freezing and thawing. J Androl 23:242–249PubMedGoogle Scholar
  9. 9.
    Calamera JC et al (2010) Effect of thawing temperature on the motility recovery of cryopreserved human spermatozoa. Fertil Steril 93:789–794PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.IVF LaboratoryUniversity Women’s HealthcareLouisvilleUSA

Personalised recommendations