Semen cryopreservation

Semen cryopreservation (commonly called sperm banking or sperm freezing) is a procedure to preserve sperm cells. Semen can be used successfully indefinitely after cryopreservation. For human sperm, the longest reported successful storage is 24 years.[1] It can be used for sperm donation where the recipient wants the treatment in a different time or place, or as a means of preserving fertility for men undergoing vasectomy or treatments that may compromise their fertility, such as chemotherapy, radiation therapy or surgery.


The most common cryoprotectant used for semen is glycerol (10% in culture medium). Often sucrose or other di-, trisaccharides are added to glycerol solution. Cryoprotectant media may be supplemented with either egg yolk or soy lecithin, with the two having no statistically significant differences compared to each other regarding motility, morphology, ability to bind to hyaluronate in vitro, or DNA integrity after thawing.[2]

Additional cryoprotectants can be used to increase sperm viability and fertility rates post-freezing. Treatment of sperm with heparin binding proteins prior to cryopreservation showed decreased cryoinjury and generation of ROS.[3] The addition of nerve growth factor as a cryoprotectant decreases sperm cell death rates and increased motility after thawing.[4] Incorporation of cholesterol into sperm cell membranes with the use of cyclodextrins prior to freezing also increases sperm viability.[5]

Semen is frozen using either a controlled-rate, slow-cooling method (slow programmable freezing or SPF) or a newer flash-freezing process known as vitrification. Vitrification gives superior post-thaw motility and cryosurvival than slow programmable freezing.[6]


Thawing at 40 °C seems to result in optimal sperm motility. On the other hand, the exact thawing temperature seems to have only minor effect on sperm viability, acrosomal status, ATP content, and DNA.[7] As with freezing, various techniques have been developed for the thawing process, both discussed by Di Santo et al.[8]


In terms of the level of sperm DNA fragmentation, up to three cycles of freezing and thawing can be performed without causing a level of risk significantly higher than following a single cycle of freezing and thawing. This is provided that samples are refrozen in their original cryoprotectant and are not going through sperm washing or other alteration in between, and provided that they are separated by density gradient centrifugation or swim-up before use in assisted reproduction technology.[9]

Effect on quality

Some evidence suggests an increase in single-strand breaks, condensation and fragmentation of DNA in sperm after cryopreservation. This can potentially increase the risk of mutations in offspring DNA. Antioxidants and the use of well-controlled cooling regimes could potentially improve outcomes.[10]

In long-term follow-up studies, no evidence has been found either of an increase in birth defects or chromosomal abnormalities in people conceived from cryopreserved sperm compared with the general population.[10]

See also


  1. Zhang, S (December 21, 2017). "A Woman Gave Birth From an Embryo Frozen for 24 Years". The Atlantic. Archived from the original on September 15, 2019. Retrieved October 13, 2019.
  2. Reed, ML; Ezeh, PC; Hamic, A; Thompson, DJ; Caperton, CL (2009). "Soy lecithin replaces egg yolk for cryopreservation of human sperm without adversely affecting postthaw motility, morphology, sperm DNA integrity, or sperm binding to hyaluronate". Fertility and Sterility. 92 (5): 1787–1790. doi:10.1016/j.fertnstert.2009.05.026. PMID 19539916.
  3. Patel, M; Gandotra, VK; Cheema, RS; Bansal, AK; Kumar, A (2016). "Seminal Plasma Heparin Binding Proteins Improve Semen Quality by Reducing Oxidative Stress during Cryopreservation of Cattle Bull Semen". Asian-Australasian Journal of Animal Sciences. 29 (9): 1247–1255. doi:10.5713/ajas.15.0586. PMC 5003984. PMID 26954172.
  4. Saeednia, S; Bahadoran, H; Amidi, F; Asadi, MH; Naji, M; Fallahi, P; Nejad, NA (2015). "Nerve growth factor in human semen: Effect of nerve growth factor on the normozoospermic men during cryopreservation process". Iranian Journal of Basic Medical Sciences. 18 (3): 292–299. doi:10.22038/IJBMS.2015.4134. PMC 4414996. PMID 25945243.
  5. Purdy, PH; Graham, JK (2004). "Effect of cholesterol-loaded cyclodextrin on the cryosurvival of bull sperm". Cryobiology. 48 (1): 36–45. doi:10.1016/j.cryobiol.2003.12.001. PMID 14969680.
  6. Vutyavanich, T; Piromlertamorn, W; Nunta, S (2010). "Rapid freezing versus slow programmable freezing of human spermatozoa". Fertility and Sterility. 93 (6): 1921–1928. doi:10.1016/j.fertnstert.2008.04.076. PMID 19243759.
  7. Calamera, JC; Buffone, MG; Doncel, GF; Brugo-Olmedo, S; de Vincentiis, S; Calamera, MM; Storey, BT; Alvarez, JG (2010). "Effect of thawing temperature on the motility recovery of cryopreserved human spermatozoa". Fertility and Sterility. 93 (3): 789–794. doi:10.1016/j.fertnstert.2008.10.021. PMID 19059590.
  8. Di Santo, M; Tarozzi, N; Nadalini, M; Borini, A (2012). "Human Sperm Cryopreservation: Update on Techniques, Effect on DNA Integrity, and Implications for ART". Advances in Urology. 2012: 854837. doi:10.1155/2012/854837. PMC 3238352. PMID 22194740.
  9. Thomson, LK; Fleming, SD; Barone, K; Zieschang, JA; Clark, AM (2010). "The effect of repeated freezing and thawing on human sperm DNA fragmentation". Fertility and Sterility. 93 (4): 1147–1156. doi:10.1016/j.fertnstert.2008.11.023. PMID 19135665.
  10. Kopeika, J.; Thornhill, A.; Khalaf, Y. (2014). "The effect of cryopreservation on the genome of gametes and embryos: principles of cryobiology and critical appraisal of the evidence". Human Reproduction Update. 21 (2): 209–227. doi:10.1093/humupd/dmu063. PMID 25519143.
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