Medium for freezing human sperm cells

SpermFreeze™ is a single step freezing medium for use with human sperm cells. The medium is clear (does not contain any milk, egg yolk, ...) and can be used direct from the bottle.
SpermFreeze™ requires 0.7ml of medium for each 1ml of semen. We also offer a more concentrated SpermFreeze™ which allows 3ml of semen to be frozen with 1ml of SpermFreeze SSP™.

SpermFreeze™ complies with the following specifications:

pH 7.2-7.9 (7.2-7.6 at release)
Sterility Sterile
Endotoxine < 0.25 EU/mL
Sperm Survival test ≥ 80% motility recovery after 4 hours exposure
Shelflife 18 months from date of produce
Albumin FDA (USA) and EMEA (Europe) compliant


Europe: CE-marked - USA: US FDA Cleared
Canada: Health Canada License - Brazil: registered

Product order codes

SPF SpermFreeze - 5x 20ml
SPF05 SpermFreeze - 25 x 5ml

The product composition can be found in the MSDS (see Resources). Additional information on some components is provided below:

Component Benefit
HEPES (20 mM)

HEPES stabilizes the pH under air (Clark and Swain 2014), therefore CO2 incubation is not required.


Cryoprotectants act by decreasing the freezing point of a substance, reducing the amount of salts and solutes present in the liquid phase of the sample and by decreasing ice formation within the spermatozoa. Importantly, the cryoprotectant should be non-toxic to the cells during the addition of the cryoprotectant and during freezing and thawing.

Glycerol is a permeating cryoprotectant which is most widely used for human sperm cryopreservation and meets all criteria indicated above. Glycerol acts on the membrane structure, permeability and stability of the lipid bilayer, the association of surface proteins and the cellular metabolism (Di Santo, et al. 2012).

Macromolecules (4 g/L HSA and sucrose(20 mM))

Cryoprotective solutions have been supplemented with macromolecules such as sucrose and human serum albumin (HSA). Several studies have shown that such molecules help to reduce physical damage and help to maintain osmotic pressure of the extracellular fluid (Shaw, et al. 2000).

Besides its cryoprotective role, HSA facilitates gamete or embryo manipulation by preventing adsorption to the surface of petri dishes and pipettes through saturation of the potential binding sites.
Also, the increased viscosity of the media, caused by the addition of HSA, promotes the ease of embryo handling and manipulation (Trounson and Gardner 2000).

Product literature

     Bandularatne E., Bongso A., Evaluation of Human Sperm Function After Repeated Freezing and Thawing, Journal of Andrology (2002),Vol.23,No.2,pp.242-249
     Bizet, P.; Saias-Magnan, J.; Jouve, E.; Grillo, JM.; Kasentry, G.; Metzler-Guillemain, C.; Perrin, J., Sperm cryopreservation before cancer treatment: a 15-year monocentric experience., Reproductive BioMedicine online (2012),Vol.24,pp.321-330
     Boitrelle, F.; Albert, M.; Theillac, C.; Ferfouri, F.; Bergere, M.; Vialard, F.; Wainer, R.; Bailly, M.; Selva, J., Cryopreservation of human spermatozoa decreases the number of motile normal spermatozoa induces nuclear vacuolization and chromatin condensation., Journal of andrology (2012),Vol.33,pp.1371-1378
     Bromage S.J., Douglas J., Falconer D.A., Lieberman B.A., Payne S.R., Factors affecting succesful outcome from ICSI in men following previous vasectomy, World Journal of Urology (2007),Vol.25,pp.519-524
     Freour, T.;Mirallie, S.; Jean, M.; Barriere, P., Sperm banking and assisted reproductive outcome in men with cancer: a 10 year's experience., Int J Clin Oncol (2012),Vol.17,No.6,pp.598-603
     Gatimel, N.;Leandri, R.; Parinaud, J. , Sperm vacuoles are not modified by freezing-thawing procedures., Reproductive Medicine online (2013),Vol.26,pp.240-246
     Konc J., Kanyo K., Cseh S., Deliveries from embryos fertilized with spermatozoa obtained from cryopreserved testicular tissue, Journal of Assisted Reproductive Genetics (2006),Vol.23,pp.247-252
     Moubasher, AE.; Din AMEES, Ali, ME.; El-Sherif, WT.; Gaber, HD., Catalase improves motility, vitality and DNA integrity of cryopreserved human spermatozoa., Andrologia (2012),Vol.45,pp.135-139
     Prisant, N.; Tubiana, R.; Lefebvre, G.; Lebray, P.; Marcelin, AG.; Thibault, V.; Rosenblum, O.; Bonmarchand, M.; Vauthier-Brouzes, D.; Golmard, JL.;Katlama, C.; Poirot, C., HIV-1 or hepatitis C chronic infection in serodiscordant infertile couples has no impact on fertility treatment outcome., Fertil. Steril. (2010),Vol.93,No.3,pp.1020-3
     Zribi, N.; Chakroun, NF.; Abdallah, B.; Elleuch, H.; Sellami, A.; Gargouri, J.; Rebai, T.; Fakhfakh, F.; Keskes, LA.;, Effect of freezing-thawing process and quercetin on human sperm survival and DNA integrity., Cryobiology (2012),Vol.65,pp.326-331

Literature concerning the components

     Clark, N.A. and Swain, J.E., Buffering systems in IVF., Culture media, Solutions, and Systems in Human ART, by P. Quinn (2014),pp.30-46
     Di Santo, M, N Tarozzi, M Nadalini, and A Borini., Human sperm cryopreservation: Update on techniques, effect on DNA integrity and implications for ART., Advances in urology (2012),Vol.12
     Shaw JM, Oranratnachai A, Trounson AO., Fundamental cryobiology of mammalian oocytes and ovarian tissue., Theriogenology (2000),Vol.53,pp.59-72
     Trounson AO, and Gardner, DK., , Handbook of in vitro Fertilization, Boca Raton, Florida: CRC Press (2000),Vol.2


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