Animal Reproduction (AR)
Animal Reproduction (AR)

Kinetic vitrification: concepts and perspectives in animal sperm cryopreservation

Bianca Barreto Barbosa; Inara Tayná Alves Evangelista; Airton Renan Bastos Soares; Danuza Leite Leão; Ricardo José Garcia Pereira; Sheyla Farhayldes Souza Domingues

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Sperm cryopreservation is an important tool for genetic diversity management programs and the conservation of endangered breeds and species. The most widely used method of sperm conservation is slow freezing, however, during the process, sperm cells suffer from cryoinjury, which reduces their viability and fertility rates. One of the alternatives to slow freezing is vitrification, that consist on rapid freezing, in which viable cells undergo glass-like solidification. This technology requires large concentrations of permeable cryoprotectants (P- CPA’s) which increase the viscosity of the medium to prevent intracellular ice formation during cooling and warming, obtaining successful results in vitrification of oocytes and embryos. Unfortunately, this technology failed when applied to vitrification of sperm due to its higher sensitivity to increasing concentrations of P-CPAs. Alternatively, a technique termed ‘kinetic sperm vitrification’ has been used and consists in a technique of permeant cryoprotectant-free cryopreservation by direct plunging of a sperm suspension into liquid nitrogen. Some of the advantages of kinetic vitrification are the speed of execution and no rate-controlled equipment required. This technique has been used successfully and with better results for motility in human (50-70% motility recovery), dog (42%), fish (82%) and donkey (21.7%). However, more studies are required to improve sperm viability after devitrification, especially when it comes to motility recovery. The objective of this review is to present the principles of kinetic vitrification, the main findings in the literature, and the perspectives for the utilization of this technique as a cryopreservation method.


assisted reproduction, cryobank, cryopreservation, cryoprotectant-free, spermatozoa


Aisen EG, Medina VH, Venturino A. Cryopreservation and post-thawed fertility of ram semen frozen in different trehalose concentrations. Theriogenology. 2002;57(7):1801-8. PMid:12041684.

Akiyama Y, Shinose M, Watanabe H, Yamada S, Kanda Y. Cryoprotectant-free cryopreservation of mammalian cells by superflash freezing. Proc Natl Acad Sci USA. 2019;116(16):7738-43. PMid:30936320.

Amann RP, Pickett BW. Principles of cryopreservation and a review of cryopreservation of stallion spermatozoa. J Equine Vet Sci. 1987;7(3):145-73.

Arav A, Yavin S, Zeron Y, Natan D, Dekel I, Gacitua H. New trends in gamete’s cryopreservation. Mol Cell Endocrinol. 2002;187(1-2):77-81. PMid:11988314.

Baiee FH, Al-Wahab A, Almusawi AA, Fitri W, Haron AW, Bustani GS. Effect of vitrification on spermatozoa quality in bull semen. Eurasian J Biosci. 2020;14:3897-904.

Barquero V, Roldan ERS, Soler C, Yániz JL, Camacho M, Valverde A. Predictive capacity of boar sperm morphometry and morphometric sub-populations on reproductive success after artificial insemination. Animals. 2021;11(4):920. PMid:33805060.

Baust JG, Gao D, Baust JM. Cryopreservation: an emerging paradigm change. Organogenesis. 2009;5(3):90-6. PMid:20046670.

Brito MM, Lúcio CF, Angrimani DS, Losano JD, Dalmazzo A, Nichi M, Vannucchi CI. Comparison of cryopreservation protocols (single and two-steps) and thawing (fast and slow) for canine sperm. Anim Biotechnol. 2017;28(1):67-73. PMid:27715465.

Caturla-Sánchez E, Sánchez-Calabuig MJ, Pérez-Gutiérrez JF, Cerdeira J, Castaño C, Santiago-Moreno J. Vitrification of dog spermatozoa: effects of two cryoprotectants (sucrose or trehalose) and two warming procedures. Cryobiology. 2018;80:126-9. PMid:29126865.

Celeghini ECC, Arruda RP, Andrade AFC, Nascimento J, Raphael CF, Rodrigues PHM. Effects that bovine sperm cryopreservation using two different extenders has on sperm membranes and chromatin. Anim Reprod Sci. 2008;104(2-4):119-31. PMid:17368970.

Cerdeira J, Sánchez-Calabuig MJ, Pérez-Gutiérrez JF, Hijon M, Castaño C, Santiago-Moreno J. Cryopreservation effects on canine sperm morphometric variables and ultrastructure: comparison between vitrification and conventional freezing. Cryobiology. 2020;95:164-70. PMid:32229272.

Colás C, Junquera C, Perez-Pe R, Cebrian-Perez JA, Muino-Blanco T. Ultrastructural study of the ability of seminal plasma proteins to protect ram spermatozoa against cold shock. Microsc Res Tech. 2009;72(8):566-72. PMid:19322897.

Consuegra C, Crespo F, Dorado J, Diaz-Jimenez M, Pereira B, Ortiz I, Hidalgo M. Vitrification of stallion sperm using 0.25 ml straws: effect of volume, concentration, and carbohydrates (sucrose/trehalose/raffinose). Anim Reprod. 2019;206:69-77. PMid:31138492.

Cuevas-Uribe R, Leibo SP, Daly J, Tiersch TR. Production of channel catfish with sperm cryopreserved by rapid non-equilibrium cooling. Cryobiology. 2011;63(3):186-97. PMid:21896271.

Curry MR, Millar JD, Watson PF. Calculated optimal cooling rates for ram and human sperm cryopreservation fail to conform with empirical observations. Biol Reprod. 1994;51(5):1014-21. PMid:7849177.

Donoghue AM, Wishart GJ. Storage of poultry semen. Anim Reprod Sci. 2000;62(1-3):201-32. PMid:10924826.

Du J, Tao J, Kleinhans FW, Mazur P, Critser JK. Water volume and osmotic behaviour of mouse spermatozoa determined by electron paramagnetic resonance. J Reprod Fertil. 1994;101(1):37-42. PMid:8064691.

Dupesh S, Rasappan, Shila, Gunasekaran K. A simple method of human sperm vitrification. MethodsX. 2019;6:2198-204.

Fácio CL, Previato LF, Machado-Paula LA, Matheus PCS, Araújo E Fo. Comparison of two sperm processing techniques for low complexity assisted fertilization: sperm washing followed by swim-up and discontinuous density gradient centrifugation. JBRA Assist Reprod. 2016;20(4):206-11. PMid:28050954.

Fahy GM, MacFarlane DR, Angell CA, Meryman HT. Vitrification as an approach to cryopreservation. Cryobiology. 1984;21(4):407-26. PMid:6467964.

Fahy GM, Rall WF. Vitrification: an overview. In: Tucker MJ, Liebermann J, editors. Vitrification in assisted reproduction. Boca Raton: CRC Press; 2007. p. 1–20.

Fahy GM, Wowk B. Principles of cryopreservation by vitrification. In: Wolkers WF, Oldenhof H, editors. Cryopreservation and freeze-drying protocols. New York: Springer-Verlag; 2015. (Methods in Molecular Biology; no. 1257).

Fahy GM. Overview of biological vitrification. In: Tucker MJ, Liebermann J, editors. Vitrification in assisted reproduction. 2nd ed. Boca Raton: CRC Press; 2015. p. 3-5.

Fahy GM. The relevance of cryoprotectant “toxicity” to cryobiology. Cryobiology. 1986;23(1):1-13. PMid:3956226.

Gao D, Critser JK. Mechanisms of cryoinjury in living cells. ILAR J. 2000;41(4):187-96. PMid:11123179.

Gautier C, Aurich C. “Fine feathers make fine birds” – The mammalian sperm plasma membrane lipid composition and effects on assisted reproduction. Anim Reprod Sci. 2022;246:106884. PMid:34776291.

Gibbons A, Bruno-Galarraga M, Fernandez J, Gonzalez-Bulnes A, Cueto M. Vitrified embryo transfer in Merino sheep under extensive conditions. Anim Reprod. 2019;16(2):297-301. PMid:33224290.

Glazar AI, Mullen SF, Liu J, Benson JD, Critser JK, Squires EL, Graham JK. Osmotic tolerance limits and membrane permeability characteristics of stallion spermatozoa treated with cholesterol. Cryobiology. 2009;59(2):201-6. PMid:19646432.

Gravance CG, Davis RO. Automated sperm morphometry analysis (ASMA) in the rabbit. J Androl. 1995;16(1):88-93. PMid:7539416.

Guthrie HD, Liu J, Critser JK. Osmotic tolerance limits and effects of cryoprotectants on motility of bovine spermatozoa. Biol Reprod. 2002;67(6):1811-6. PMid:12444057.

Hagiwara M, Choi JH, Devireddy RV, Roberts KP, Wolkers WF, Makhlouf A, Bischof JC. Cellular biophysics during freezing of rat and mouse sperm predicts post-thaw motility. Biol Reprod. 2009;81(4):700-6. PMid:19535788.

Hammerstedt RH, Keith AD, Snipes W, Amann RP, Arruda D, Griel LC Jr. Use of spin labels to evaluate effects of cold shock and osmolality on sperm. Biol Reprod. 1978;18(4):686-96. PMid:656535.

Hidalgo M, Diaz-Jimenez M, Consuegra C, Pereira B, Dorado J. Vitrification of donkey sperm: is it better using permeable cryoprotectants? Animals. 2020;10(9):1462. PMid:32825370.

Hidalgo M. Recent advances in donkey sperm vitrification. Reprod Domest Anim. 2021;56(10):1274-8. PMid:34418185.

Hoagland H, Pincus G. Revival of mammalian sperm after immersion in liquid nitrogen. J Gen Physiol. 1942;25(3):337-44. PMid:19873277.

Holt WV. Basic aspects of frozen storage of semen. Anim Reprod Sci. 2000;62(1-3):3-22. PMid:10924818.

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. PMid:12675999.

Isachenko E, Isachenko V, Katkov II, Rahimi G, Schöndorf T, Mallmann P, Dessole S, Nawroth F. DNA integrity and motility of human spermatozoa after standard slow freezing versus cryoprotectant-free vitrification. Hum Reprod. 2004a;19(4):932-9. PMid:15016773.

Isachenko E, Isachenko V, Weiss JM, Kreienberg R, Katkov II, Schulz M, Lulat A, Risopatrón MJ, Sánchez R. Acrosomal status and mitochondrial activity of human spermatozoa vitrified with sucrose. Reproduction. 2008;136(2):167-73. PMid:18483075.

Isachenko V, Isachenko E, Katkov II, Montag M, Dessole S, Nawroth F, Van Der Ven H. Cryoprotectant-free cryopreservation of human spermatozoa by vitrification and freezing in vapor: effect on motility, DNA integrity, and fertilization ability. Biol Reprod. 2004b;71(4):1167-73. PMid:15175233.

Isachenko V, Isachenko E, Montag M, Zaeva V, Krivokharchenko I, Nawroth F, Dessole S, Katkov II, Van der Ven H. Clean technique for cryoprotectant-free vitrification of human spermatozoa. Reprod Biomed Online. 2005;10(3):350-4. PMid:15820041.

Isachenko V, Isachenko E, Petrunkina AM, Sanchez R. Human spermatozoa vitrified in the absence of permeable cryoprotectants: birth of two healthy babies. Reprod Fertil Dev. 2012b;24(2):323-6. PMid:22281078.

Isachenko V, Isachenko E, Petrunkina AM, Sánchez R. Human spermatozoa vitrified in the absence of permeable cryoprotectants: birth of two healthy babies. Reprod Fertil Dev. 2012;24(2):323-6. PMid:22281078.

Isachenko V, Maettner R, Petrunkina AM, Mallmann P, Rahimi G, Sterzik K, Sánchez R, Risopatron J, Damjanoski I, Isachenko E. Cryoprotectant-free vitrification of human spermatozoa in large (up to 0.5 mL) volume: a novel technology. Clin Lab. 2011;57(9-10):643-50. PMid:22029178.

Isachenko V, Maettner R, Petrunkina AM, Sterzik K, Mallmann P, Rahimi G, Sanchez R, Risopatron J, Damjanoski I, Isachenko E. Vitrification of human ICSI/IVF spermatozoa without cryoprotectants: new capillary technology. J Androl. 2012a;33(3):462-8. PMid:21719694.

Isachenko V, Rahimi G, Mallmann P, Sánchez R, Isachenko E. Technologies of cryoprotectant-free vitrification of human spermatozoa: asepticity as criterion of effectiveness. Andrology. 2017;5(6):1055-63. PMid:28992376.

Isachenko V, Sánchez R, Rahimi G, Mallmann P, Isachenko E, Merzenich M. Cryoprotectant‐free vitrification of spermatozoa: fish as a model of human. Andrologia. 2019;51(1):e13166. PMid:30298523.

Jang TH, Park SC, Yang JH, Kim JY, Seok JH, Park US, Choi CW, Lee SR, Han J. Cryopreservation and its clinical applications. Integr Med Res. 2017;6(1):12-8. PMid:28462139.

Karlsson JO, Cravalho EG, Borel Rinkes IH, Tompkins RG, Yarmush ML, Toner M. Nucleation and growth of ice crystals inside cultured hepatocytes during freezing in the presence of dimethyl sulfoxide. Biophys J. 1993;65(6):2524-36. PMid:8312489.

Katkov II, Bolyukh VF, Chernetsov OA, Dudin PI, Grigoriev AY, Isachenko V, Isachenko E, Lulat AG-M, Moskovtsev SI, Petrushko MP, Pinyaev VI, Sokol KM, Sokol YI, Sushko AB, Yakhnenko I. Kinetic vitrification of spermatozoa of vertebrates: what can we learn from nature? In: Katkov I , editor. Current frontiers in cryobiology. Croatia: IntechOpen; 2012. p. 3-40.

Khalil WA, El-Harairy MA, Zeidan AEB, Hassan MAE, Mohey-Elsaeed O. Evaluation of bull spermatozoa during and after cryopreservation: structural and ultrastructural insights. Int J Vet Sci Med. 2018;6(Suppl 1):S549-56. PMid:30761321.

Kim S, Lee Y, Yang H, Kim Y. Rapid freezing without cooling equilibration in canine sperm. Anim Reprod Sci. 2012;130(1-2):111. PMid:22277841.

Kleinhans FW, Travis VS, Du J, Villines PM, Colvin KE, Critser JK. Measurement of human sperm intracellular water volume by electron spin resonance. J Androl. 1992;13(6):498-506. PMid:1338068.

Kuleshova LL, Lopata A. Vitrification can be more favorable than slow cooling. Fertil Steril. 2002;78(3):449-54. PMid:12215314.

Li Y, Zhou L, Lv M, Ge P, Liu Y, Zhou D. Vitrification and conventional freezing methods in sperm cryopreservation: a systematic review and meta-analysis. Eur J Obstet Gynecol Reprod Biol. 2019;233:84-92. PMid:30580229.

Luyet B, Rapatz G. Patterns of ice formation in some aqueous solutions. Biodynamica. 1958;8:1-68.

Luyet B. The vitrification of organic colloids and protoplasm. Biodynamica. 1937a;1:1-14.

Luyet B. Working hypotheses on the nature of life. Biodynamica. 1937b;1:1-7.

Luyet BJ, Hodapp EL. Revival of Frog’s Spermatozoa Vitrified in Liquid Air. Exp Biol Med (Maywood). 1938;39(3):433-4.

MacFarlane DR. Devitrification in glass-forming aqueous solutions. Cryobiology. 1986;23(3):230-44.

Maroto-Morales A, Ramón M, García-Alvarez O, Soler AJ, Esteso MC, Martínez-Pastor F, Pérez-Guzmán MD, Garde JJ. Characterization of ram (Ovis aries) sperm head morphometry using the Sperm-Class Analyzer. Theriogenology. 2010;73(4):437-48. PMid:20018357.

Mazur P, Katkov II, Katkova N, Critser JK. The enhancement of the ability of mouse sperm to survive freezing and thawing by the use of high concentrations of glycerol and the presence of an E. coli membrane preparation OxyraseTM to lower the oxygen concentration. Cryobiology. 2000;40(3):187-209. PMid:10860619.

Mazur P. Principles of cryobiology. In: Fuller BJ, Lane NL, Benson EE, editors. Life in the frozen state. Boca Raton: CRC Press LLC; 2004. p. 3-65.

Medeiros CM, Forell F, Oliveira AT, Rodrigues JL. Current status of sperm cryopreservation: why isn’t it better? Theriogenology. 2002;57(1):327-44. PMid:11775978.

Merino O, Risopatrón J, Sánchez R, Isachenko E, Figueroa E, Valdebenito I, Isachenko V. Fish (Oncorhynchus mykiss) spermatozoa cryoprotectant-free vitrification: stability of mitochondrion as criterion of effectiveness. Anim Reprod Sci. 2011;124(1-2):125-31. PMid:21392903.

Meryman HT. Cryoprotective agentes. Cryobiology. 1971;8(2):173-83. PMid:5578883.

Mocé E, Fajardo AJ, Graham JK. Human sperm cryopreservation. Eur Med J. 2016;1:86-91.

Mosca F, Madeddu M, Sayed AA, Zaniboni L, Iaffaldano N, Cerolini S. Combined effect of permeant and non-permeant cryoprotectants on the quality of frozen/thawed chicken sperm. Cryobiology. 2016;73(3):343-7. PMid:27717656.

Mphaphathi ML, Luseba D, Sutherland B, Nedambale TL. Comparison of slow freezing and vitrification methods for Venda cockerel’s spermatozoa. Open J Anim Sci. 2012;3(3):204-10.

Nagashima JB, Sylvester SR, Nelson JL, Cheong SH, Mukai C, Lambo C, Flanders JA, Meyers-Wallen VN, Songsasen N, Travis AJ. Live Births from Domestic Dog (Canis familiaris) Embryos Produced by In Vitro Fertilization. PLoS One. 2015;10(12):1.

Nagata MPB, Egashira J, Katafuchi N, Endo K, Ogata K, Yamanaka K, Yamanouchi T, Matsuda H, Hashiyada Y, Yamashita K. Bovine sperm selection procedure prior to cryopreservation for improvement of postthawed semen quality and fertility. J Anim Sci Biotechnol. 2019;10(1):91. PMid:31807306.

Nawroth F, Isachenko V, Dessole S, Rahimi G, Farina M, Vargiu N, Mallmann P, Dattena M, Capobianco G, Peters D, Orth I, Isachenko E. Vitrification of human spermatozoa without cryoprotectants. Cryo Lett. 2002;23(2):93-102. PMid:12050777.

Nunes JF. Inseminação artificial em caprinos. In: Gonsalvez PBD, Figueiredo JR, Freitas VJF, editors. Biotécnicas aplicadas à reprodução animal. São Paulo: Varela; 2002. p. 111-25.

O’Connell M, McClure N, Powell LA, Steele EK, Lewis SEM. Differences in mitochondrial and nuclear DNA status of high-density and low-density sperm fractions after density centrifugation preparation. Fertil Steril. 2003;79(Suppl 1):754-62. PMid:12620488.

O’Neill HC, Nikoloska M, Ho H, Doshi A, Maalouf W. Improved cryopreservation of spermatozoa using vitrification: comparison of cryoprotectants and a novel device for long-term storage. J Assist Reprod Genet. 2019;36(8):1713-20. PMid:31273587.

Ozkavukcu S, Erdemli E, Isik A, Oztuna D, Karahuseyinoglu S. Effects of cryopreservation on sperm parameters and ultrastructural morphology of human spermatozoa. J Assist Reprod Genet. 2008;25(8):403-11. PMid:18704674.

Ozkavukcu S, Erdemli E. Cryopreservation: basic knowledge and biophysical effects. J Ankara Med Sch. 2002;24:187-96.

Pan C, Yu S, Zhang P, Wang B, Zhu Z, Liu Y, Zeng W. Effect of sucrose on cryopreservation of pig spermatogonial stem cells. J Integr Agric. 2017;16(5):1120-9.

Paoli D, Lombardo F, Lenzi A, Gandini L. Sperm cryopreservation: effects on chromatin structure. In: Baldi E, Muratori M, editors. Genetic damage in human spermatozoa. Switzerland: Springer; 2014. p. 137-50. (Advances in Experimental Medicine and Biology; no. 791).

Pegg DE. Principles of cryopreservation. In: Wolkers WF, Oldenhof H, editors. Cryopreservation and freeze-drying protocols. 3rd ed. New York: Springer; 2015. p. 3-19. (Methods in Molecular Biology; no. 1257).

Pegg DE. Principles of cryopreservation. Methods Mol Biol. 2007;368:39-57. PMid:18080461.

Pérez-Sánchez F, de Monserrat JJ, Soler C. Morphometric analysis of human sperm morphology. Int J Androl. 1994;17(5):248-55. PMid:7698850.

Phillips PH, Lardy HA. A yolk buffer pabulum for the preservation of bull semen. J Dairy Sci. 1940;23(5):399-404.

Polge C, Smith A, Parkes A. Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature. 1949;164(4172):666. PMid:18143360.

Rall WF. Factors affecting the survival of mouse embryos cryopreserved by vitrification. Cryobiology. 1987;24(5):387-402. PMid:3652721.

Rosato MP, Iaffaldano N. Cryopreservation of rabbit semen: comparing the effects of different cryoprotectants, cryoprotectant-free vitrification, and the use of albumin plus osmoprotectants on sperm survival and fertility after standard vapor freezing and vitrification. Theriogenology. 2013;79(3):508-16. PMid:23218394.

Sánchez MV, Bastir M, Roldan ERS. Geometric morphometrics of rodent sperm head shape. PLoS One. 2013;8(11):1-10. PMid:24312234.

Sánchez R, Risopatrón J, Schulz M, Villegas J, Isachenko V, Kreinberg R, Isachenko E. Canine sperm vitrification with sucrose: effect on sperm function. Andrologia. 2011;43(4):233-41. PMid:21486402.

Santo MD, 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. PMid:22194740.

Schiewe MC, Mullen SF. Vitrification: fundamental principles and its application for cryopreservation of human reproductive cells. In: Bozkurt Y, editor. Cryopreservation biotechnology in biomedical and biological sciences. London: IntechOpen; 2018.

Schulz M, Risopatrón J, Uribe P, Isachenko E, Isachenko V, Sánchez R. Human sperm vitrification: a scientific report. Andrology. 2020;8(6):1642-50. PMid:32598551.

Shaffner CS, Henderson EW, Card CG. Viability of spermatozoa of the chicken under various environmental conditions. Poult Sci. 1941;20(3):259-65.

Shaw JM, Jones GM. Terminology associated with vitrification and other cryopreservation procedures for oocytes and embryos. Hum Reprod Update. 2003;9(6):583-605. PMid:14714593.

Shaw JM, Kuleshova LL, Macfarlane DR, Trounson AO. Vitrification properties of solutions of ethylene glycol in saline containing PVP, Ficoll, or Dextran. Cryobiology. 1997;35(3):219-29. PMid:9367610.

Sherman JK. Cryopreservation of human semen. In: Keel BA, Webster BW, editors. CRC handbook of the laboratory diagnosis and treatment of infertility. Boca Raton: CRC Press; 1990. p. 229-59.

Shi X, Wang T, Qiu ZL, Li K, Li L, Chan CPS, Chan SM, Li T, Quan S. Effects of mechanical stresses on sperm function and fertilization rate in mice. Syst Biol Reprod Med. 2016;62(2):152-9. PMid:26889695.

Sieme H, Oldenhof H, Wolkers WF. Mode of action of cryoprotectants for sperm preservation. Anim Reprod Sci. 2016;169:2-5. PMid:26936658.

Silva AR, Lima GL, Peixoto G, Souza ALP. Cryopreservation in mammalian conservation biology: current applications and potential utility. Research and Reports in Biodiversity Studies. 2015;4:1-8.

Silva HVR, Silva AR, Silvada LDM, Comizzoli P. Sperm cryopreservation and banking for the conservation of neotropical carnivores. Biopreserv Biobank. 2019;17(2):183-8. PMid:30589562.

Slabbert M, du Plessis SS, Huyser C. Large volume cryoprotectant-free vitrification: an alternative to conventional cryopreservation for human spermatozoa. Andrologia. 2015;47(5):594-9. PMid:25040423.

Smith AU, Polge C. Survival of spermatozoa at low temperatures. Nature. 1950;166(4225):668-9. PMid:14780190.

Sun W, Jiang S, Su J, Zhang J, Bao X, Ding R, Shi P, Li S, Wu C, Zhao G, Cao G, Sun QY, Yu H, Li X. The effects of cryopreservation on the acrosome structure, enzyme activity, motility, and fertility of bovine, ovine, and goat sperm. Anim Reprod. 2020;17(4):e20200219. PMid:34084227.

Swain JE, Smith GD. Cryoprotectants. In: Chian RC, Quinn P, editors. Fertility Cryopreservation. Cambridge: Cambridge University Press; 2010. p. 24-38.

Swanson W, Bateman H, Vansandt L. Urethral catheterization and sperm vitrification for simplified semen banking in felids. Reprod Domest Anim. 2017;52(Suppl 2):255-60. PMid:27807883.

Tani M, Neely JR. Role of intracellular Na+ in Ca2+ overload and depressed recovery of ventricular function of reperfused ischemic rat hearts. Possible involvement of H+-Na+ and Na+-Ca2+ exchange. Circ Res. 1989;65(4):1045-56. PMid:2551525.

Tao Y, Sanger E, Saewu A, Saewu A, Leveille M. Human sperm vitrification: the state of the art. Reprod Biol Endocrinol. 2020;18(1):17. PMid:32145746.

Thachil JV, Jewett MA. Preservation techniques for human semen. Fertil Steril. 1981;35(5):546-8. PMid:7227570.

Thananurak P, Chuaychu-noo N, Thélie A, Phasuk Y, Vongpralub T, Blesbois E. Sucrose increases the quality and fertilizing ability of cryopreserved chicken sperms in contrast to rafinose. Poult Sci. 2019;98(9):4161-71. PMid:31065720.

Tomlinson MJ, Moffatt O, Manicardi GC, Bizzaro D, Afnan M, Sakkas D. Interrelationships between seminal parameters and sperm nuclear DNA damage before and after density gradient centrifugation: implications for assisted conception. Hum Reprod. 2001;16(10):2160-5. PMid:11574509.

Valojerdi MR, Eftekhari-Yazdi P, Karimian L, Hassani F, Movaghar B. Vitrification versus slow freezing gives excellent survival, post warming embryo morphology and pregnancy outcomes for human cleaved embryos. J Assist Reprod Genet. 2009;26(6):347-54. PMid:19513822.

Valverde A, Arenán H, Sancho M, Contell J, Yániz J, Fernández A, Soler C. Morphometry and subpopulation structure of Holstein bull spermatozoa: variations in ejaculates and cryopreservation straws. Asian J Androl. 2016;18(6):851-7. PMid:27678464.

Varago FC, Saliba WP, Alvim MTT, Vasconcelos AB, Oliveira CH, Stahlberg R, Lagares MA. Vitrification of in vitro produced Zebu embryos. Anim Reprod. 2006;3(3):353-8.

Villaverde-Morcillo S, Esteso MC, Castaño C, Díaz AT, López-Sebastián A, Campo JL, Santiago-Moreno J. Influence of Staining Method on the Values of Avian Sperm Head Morphometric Variables. Reprod Domest Anim. 2015;50(5):750-5. PMid:26192019.

Wang M, Todorov P, Wang W, Isachenko E, Rahimi G, Mallmann P, Isachenko V. Cryoprotectants-free vitrification and conventional freezing of human spermatozoa: a comparative transcript profiling. Int J Mol Sci. 2022;23(6):3047.

World Health Organization – WHO. Laboratory manual for the examination and processing of human semen. 5. Geneva: WHO Press; 2010.

Wowk B. Thermodynamic aspects of vitrification. Cryobiology. 2010;60(1):11-22. PMid:19538955.

Xue X, Wang W, Shi J, Zhang L, Zhao W, Shi W, Guo B, Qin Z. Efficacy of swim-up versus density gradient centrifugation in improving sperm deformity rate and DNA fragmentation index in semen samples from teratozoospermic patients. J Assist Reprod Genet. 2014;31(9):1161-6. PMid:25015033.

Yashaswi S, Mona S. Biophysics of Cryopreservation. Int J Thermodyn. 2022;25(1):17-27.

Zainuddin ZZ, Tarmizi MRM, Yap KC, Comizzoli P, Symphorosa S. First evaluations and cryopreservation of semen samples from Sunda Clouded Leopards (Neofelis diardi). Animals. 2020;10(6):1072.

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