Stallion sperm cryopreservation: concepts in cryopreservation-induced sperm damage, processing, analysis, and utilization in assisted reproductive technologies

  • Camilo Hernández-Avilés Equine Fertility Laboratory, Department of Large Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USA
Keywords: Stallion sperm, cryopreservation, cryoprotectants, semen extender, postthaw quality

Abstract

Semen cryopreservation remains the only viable method for long-term stallion sperm preservation. Increased demand for frozen/thawed sperm in Assisted Reproductive Technologies (ARTs) continues to result in a considerable body of research determining the effects of cryopreservation on stallion sperm and clinical interpretation of sperm quality in frozen/thawed semen. Some concepts regarding the impacts of cryopreservation on stallion sperm or methods for sperm cryopreservation might limit the widespread implementation of this technique in private practice settings. Clinically relevant data on stallion sperm cryopreservation, various protocols for sperm cryopreservation, concepts on postthaw sperm quality analysis, and utilization of frozen/thawed sperm for ARTs are summarized.

Downloads

Download data is not yet available.

References


1.
Iljinskaja T: The effect of various factors on stallion spermatozoa frozen to –70°C. Anim Breed Abst 1957;26:132.


2.
Polge C, Minotakis C: Deep-freezing of jackass and stallion semen. Proc Int Cong Anim Reprod Artif Insem 1964;7:545.


3.
Nagase H, Soejima S, Niwa T, et al: Studies on the freezing storage of stallion semen. I. Fertility results of stallion frozen semen in concentrated pellet form. Jpn J Anim Reprod 1966;2:48-51. doi: 10.1262/jrd1955.12.48


4.
Nishikawa Y: Studies on the preservation of raw and frozen horse semen. J Reprod Fertil 1975;23:99-104.


5.
Krause D, Grove D: Deep-freezing of jackass and stallion semen in concentrated pellet form. J Reprod Fertil 1967;14:139-141. doi: 10.1530/jrf.0.0140139


6.
Bielański W, Bilik K, Zapletal Z: Artificial insemination in horses. II. Preliminary trials with freezing semen in liquid nitrogen (–196°C). Med Wet 1969;25:287-289.


7.
Barker CAV, Gandier JCC: Pregnancy in a mare resulting from frozen epididymal spermatozoa. Can J Comp Med Vet Sci 1957;21:47-51.


8.
Pickett BW, Amann RP: Cryopreservation of semen. In: McKinnon AO, Voss JL: editors. Equine Reproduction. Philadelphia, Lea & Febiger; 1993. p. 769-789.


9.
Klug E, Treu H, Hillmann H, et al: Results of insemination of mares with fresh and frozen stallion semen. J Reprod Fertil 1975;23:107-110.


10.
Müller Z: Practicalities of insemination of mares with deep-frozen semen. J Reprod Fertil 1987;35:121-125.


11.
Vidament M: French field results (1985–2005) on factors affecting fertility of frozen stallion semen. Anim Reprod Sci 2005;89:115-136. doi: 10.1016/j.anireprodsci.2005.07.003


12.
Sieme H, Schäfer T, Stout TAE, et al: The effects of different insemination regimes on fertility in mares. Theriogenology 2003;60:1153-1164. doi: 10.1016/S0093-691X(03)00113-4


13.
Reger HP, Bruemmer JE, Squires EL, et al: Effects of timing and placement of cryopreserved semen on fertility of mares. Equine Vet Educ 2003;15:101-106. doi: 10.1111/j.2042-3292.2003.tb00224.x


14.
Loomis PR, Squires EL: Frozen semen management in equine breeding programs. Theriogenology 2005;64:480-491. doi: 10.1016/j.theriogenology.2005.05.028


15.
Miller CD: Optimizing the use of frozen-thawed equine semen. Theriogenology 2008;70:463-468. doi: 10.1016/j.theriogenology.2008.04.037


16.
Immonen I, Cuervo-Arango J: Effect of timing of postovulatory insemination relative to human chorionic gonadotrophin/buserelin treatment with 1 straw of frozen-thawed semen on mare fertility. J Equine Vet Sci 2020;87:102900. doi: 10.1016/j.jevs.2019.102900


17.
Loomis PR. The equine frozen semen industry. Anim Reprod Sci 2001;68:191-200. doi: 10.1016/S0378-4320(01)00156-7


18.
Samper JC, Morris CA: Current methods for stallion sperm cryopreservation: a survey. Theriogenology 1998;49:895-903. doi: 10.1016/S0093-691X(98)00039-9


19.
Loomis PR, Graham JK: Commercial semen freezing: individual male variation in cryosurvival and the response of stallion sperm to customized freezing protocols. Anim Reprod Sci 2008;105:119-128. doi: 10.1016/j.anireprosci.2007.11.010


20.
Samper JC, Hellander JC, Crabo BG: Relationship between the fertility of fresh and frozen stallion semen and semen quality. J Reprod Fertil 1991;44:107-114.


21.
Wilhelm KM, Graham JK, Squires EL: Comparison of the fertility of cryopreserved stallion spermatozoa with sperm motion analyses, flow cytometric evaluation, and zona-free hamster oocyte penetration. Theriogenology 1996;46:559-578. doi: 10.1016/0093-691X(96)00209-9


22.
Kirk ES, Squires EL, Graham JK: Comparison of in vitro laboratory analyses with the fertility of cryopreserved stallion spermatozoa. Theriogenology 2005;64:1422–1439. doi: 10.1016/j.theriogenology.2005.03.006


23.
Kuisma P, Andersson M, Koskinen E, et al: Fertility of frozen-thawed stallion semen cannot be predicted by the current laboratory methods. Acta Vet Scand 2006;48:14. doi: 10.1186/1751-0147-48-14


24.
Hammerstedt RH, Graham JK, Nolan JP: Cryopreservation of mammalian sperm: what we ask them to survive. J Androl 1990;11:73-88. doi: 10.1002/j.1939-4640.1990.tb01583.x


25.
Graham JK, Kunze E, Hammerstedt RH: Analysis of sperm cell viability, acrosomal integrity, and mitochondrial function using flow cytometry. Biol Reprod 1990;43:55-64. doi: 10.1095/biolreprod43.1.55


26.
Graham JK, Nolan JP, Hammerstedt RH: Effect of dilaurophosphatidylcholine liposomes on motility, induction of the acrosome reaction, and subsequent egg penetration of ram epididymal sperm. Biol Reprod 1991;44:1092-1099. doi: 10.1095/biolreprod44.6.1092


27.
Nolan JP, Graham JK, Hammerstedt RH: Artificial induction of exocytosis in bull sperm. Arch Biochem Biophys 1992;292:311-322. doi: 10.1016/0003-9861(92)90084-a


28.
Singer SL, Nicolson GL: The fluid mosaic model of the structure of cell membranes. Science 1972;175:720-731. doi: 10.1126/science.175.4023.720


29.
Parks JE, Lynch DV: Lipid composition and themotropic phase behavior of boar, bull, stallion, and rooster sperm membranes. Cryobiology 1992;29:255-266. doi: 10.106/0011-2240(92)90024-v


30.
Tapia JA, Macias-Garcia B, Miro-Morán A, et al: The membrane of the mammalian spermatozoa: much more than an inert envelope. Reprod Dom Anim 2012;47:65-75. doi: 10.1111/j.1439-0531.2012.02046.x


31.
Langlais J, Roberts KD: A molecular membrane model of sperm capacitation and the acrosome reaction of mammalian spermatozoa. Gamete Res 1985;12:183-224. doi: 10.1002/mrd.1120120209


32.
Nolan JP, Hammerstedt RH: Regulation of membrane stability and the acrosome reaction in mammalian sperm. FASEB J 1997;11 670-682. doi: 10.1096/fasebj.11.8.9240968


33.
Stein KK, Primakoff P, Myles D: Sperm-egg fusion: events at the plasma membrane. J Cell Sci 2004;117:6269-6274. doi: 10.1242/jcs.01598


34.
Gadella BM, Luna C: Cell biology and functional dynamics of the mammalian sperm surface. Theriogenology 2014;81:74-84. doi: 10.1016/j.theriogenology.2013.09.005


35.
Graham JK: Principles of cooled semen. In: McKinnon AO, Squires EL, Vaala WE, et al. Equine Reproduction, 2nd edition. Ames; Wiley-Blackwell: 2011. p. 1308-1315.


36.
Hoffmann N, Oldenhof H, Morandini C, et al: Optimal concentrations of cryoprotective agents for semen from stallions that are classified ‘good’ or ‘poor’ for freezing. Anim Reprod Sci 2011;125:112-118. doi: 10.1016/j.anireprosci.2011.03.001


37.
Holt WV, North RD: Partially irreversible cold-induced lipid phase transitions in mammalian sperm plasma membrane domains: freeze-fracture study. J Exp Zool 1984;230:473-483. doi: 10.1002/jez.1402300316


38.
De Leeuw FE, Chen HC, Colenbrander B, et al: Cold-induced ultrastructural changes in bull and boar sperm plasma membranes. Cryobiology 1990;27:171-183. doi: 10.1016/0011-2240(90)90009-s


39.
Mantsch HH, McElhaney RN: Phospholipid phase transitions in model and biological membranes as studied by infrared spectroscopy. Chem Phys Lipids 1991;57:213-226. doi: 10.1016/0009-3084(91)90077-O


40.
Oldenhof H, Friedel K, Akhoondi M, et al: Membrane phase behavior during cooling of stallion sperm and its correlation with freezability. Mol Membr Biol 2012;29:95-106. doi: 10.3109/09687688.2012.674161


41.
Drobnis EZ, Crowe LM, Berger T, et al: Cold shock damage is due to lipid phase transitions in cell membranes: A demonstration using sperm as a model. J Exp Zool 1993;265:432-437. doi: 10.1002/jez.1402650413


42.
Oldenhof H, Gojowsky M, Wang S, et al: Osmotic stress and membrane phase changes during freezing of stallion sperm: mode of action of cryoprotective agents. Biol Reprod 2013;88:68. doi: 10.1095/biolreprod.112.104661


43.
Mazur P: Freezing of living cells: mechanisms and implications. Am J Physiol 1984;247: 125-142. doi: 10.1152/ajpcell.1984.247.3.C125


44.
Ball BA, Vo A: Osmotic tolerance of equine spermatozoa and the effects of soluble cryoprotectants on equine sperm motility, viability, and mitochondrial membrane potential. J Androl 2001;22:1061-1069. doi: 10.1002/j.1939-4640.2001.tb03446.x


45.
Pommer AC, Rutllant J, Meyers SA: The role of osmotic resistance on equine spermatozoal function. Theriogenology 2002;58:1373-1384. doi: 10.1016/s0093-691x(02)01039-7


46.
Glazar AI, Mullen SF, Liu J, et al: Osmotic tolerance limits and membrane permeability characteristics of stallion spermatozoa treated with cholesterol. Cryobiology 2009;59:201-206. doi: 10.1016/j.cryobiol.2009.07.009


47.
González-Fernández L, Morrell JM, Peña FJ, et al: Osmotic shock induces structural damage on equine spermatozoa plasmalemma and mitochondria. Theriogenology 2012;78:415-422. doi: 10.1016/j.theriogenology.2012.02.021


48.
Wessel MT, Ball BA: Step-wise dilution for removal of glycerol from fresh and cryopreserved equine spermatozoa. Anim Reprod Sci 2004;84:147-156. doi: 10.1016/j.anireprosci.2003.12.004


49.
Macías García B, Miró Morán A, González Fernández L, et al: The mitochondria of stallion spermatozoa are more sensitive than the plasmalemma to osmotic-induced stress: role of c-Jun N-terminal kinase (JNK) pathway. J Androl 2012;33:105-113. doi: 10.2164/jandrol.110.011957


50.
Ortega-Ferrusola C, Sotillo-Galán Y, Varela-Fernández E, et al: Detection of apoptosis like changes during the cryopreservation process in equine sperm. J Androl 2008;29:213-221. doi: 10.2164/j.androl.107.003640


51.
Ortega-Ferrusola C, González-Fernández L, Macías García B, et al: Lipid peroxidation, assessed with BODIPY-C11 increases after cryopreservation of stallion spermatozoa, is stallion dependent, and relates to “apoptosis like” changes. Reproduction 2009;138:55-63. doi: 10.153/REP-08-0484


52.
Ortega-Ferrusola C, González-Fernández L, Macías García B, et al: Inhibition of the mitocondrial permeability transition pore reduces “apoptosis like changes” during cryopreservation of stallion spermatozoa. Theriogenology 2010;74:458-465. doi: 10.1016/j.theriogenology.2010.02.029


53.
Martín Muñoz P, Ortega Ferrusola C, Vizuete G, et al: Depletion of intracellular thiols and increased production of 4-hydroxynonenal that occur during cryopreservation of stallion spermatozoa lead to caspase activation, loss of motility, and cell death. Biol Reprod 2015;93:143. doi: 10.1095/biolreprod.115.132878


54.
Ortega Ferrusola C, Anel-López L, Ortiz-Rodríguez JM, et al: Stallion spermatozoa surviving freezing and thawing experience membrane depolarization and increased intracellular Na+. Andrology 2017;5:1174-1182. doi: 10.1111/andr.12419


55.
Burnaugh L, Ball BA, Sabeur K, et al: Osmotic stress stimulates generation of superoxide anion by spermatozoa in horses. Anim Reprod Sci 2010;117:249-260. doi: 10.1016/j.anireprosci.2009.05.014


56.
Peña FJ, Plaza Davila M, Ball BA, et al: The impact of reproductive technologies on stallion mitochondrial function. Reprod Dom Anim 2015;50:529-537. doi: 10.1111/rda.12551


57.
Peña FJ, O’Flaherty C, Ortíz Rodríguez JM, et al: Redox regulation and oxidative stress: the particular case of the stallion spermatozoa. Antioxidants 2019;8:567. doi: 10.3390/antiox8110567


58.
Peña FJ, Gibb Z: Oxidative stress and the long-term storage of horse spermatozoa. Reproduction 2022;164:135-144. doi: 10.1530/REP-22-0264


59.
Gibb Z, Lambourne SR, Aitken RJ: The paradoxical relationship between stallion fertility and oxidative stress. Biol Reprod 2014;91:77. doi: 10.1095/biolreprod.114.118539


60.
Plaza Davila M, Martin Muñoz P, Gallardo Bolaños JM, et al: Mitochondrial ATP is required for the maintenance of membrane integrity in stallion spermatozoa, whereas motility requires both glycolysis and oxidative phosphorylation. Reproduction 2016;152:683-694. doi: 10.1530/REP-16-0409


61.
Darr CR, Varner DD, Teague SR, et al: Lactate and pyruvate are major sources of energy for stallion sperm with dose effects on mitochondrial function, motility, and ROS production. Biol Reprod 2016;95:34. doi: 10.1095/biolreprod.116.140707


62.
Ramírez-Agámez L, Hernández-Avilés C, Ortíz I, et al: Lactate as the sole energy substrate induces spontaneous acrosome reaction in viable stallion spermatozoa. Andrology 2024;12:459-471. doi: 10.1111/andr.13479


63.
Martín-Cano FM, Gaitskell-Phillips G, Ortíz-Rodríguez JM, et al: Proteomic profiling of stallion spermatozoa suggests changes in sperm metabolism and compromised redox regulation after cryopreservation. J Proteom 2020;221:103765. doi: 10.1016/j.prot.2020.103765


64.
Gaitskell-Phillips G, Martín-Cano FM, Ortíz-Rodríguez JM, et al: In stallion spermatozoa, superoxide dismutase (Cu-Zn) (SOD1) and the aldo-keto-reductase family 1 member (AKR1B1) are the proteins most significantly reduced by cryopreservation. J Proteome Res 2021;20:2435-2446. doi: 10.1021/acs.jproteome.0c00932


65.
Gaitskell-Phillips G, Martín-Cano FM, Ortíz-Rodríguez JM, et al: Differences in the proteome of stallion spermatozoa explain stallion-to-stallion variability in sperm quality post thaw. Biol Reprod 2021;104:1097-1113. doi: 10.1093/biolre/ioab003


66.
Gaitskell-Phillips G, Martín-Cano FM, Ortiz-Rodríguez JM, et al: The seminal plasma proteins Peptidyl arginine deaminase 2, rRNA adenine N (6)-methyltransferase and KIAA0825 are linked to better motility post thaw in stallions. Theriogenology 2022;177:34–41. doi: 10.1016/j.theriogenology.2021.10.010


67.
Darin-Bennett A, White IG: Influence of cholesterol content of mammalian spermatozoa to susceptibility to cold-shock. Cryobiology 1977;14:466-470. doi: 10.1016/0011-2240(77)90008-6


68.
Moran DM, Jasko DJ, Squires EL, et al: Determination of temperature and cooling rate which induce cold shock in stallion spermatozoa. Theriogenology 1992;38:999-1012. doi: 10.1016/0093-691x(92)90114-7


69.
Heitland AV, Jasko DJ, Squires EL, et al: Factors affecting motion characteristics of frozen-thawed stallion spermatozoa. Equine Vet J 1996;28:47-53. doi: 10.1111/j.2042-3306.1996.tb01589.x


70.
Salazar Jr., JL, Teague SR, Love CC, et al: Effect of cryopreservation protocol on postthaw characteristics of stallion sperm. Theriogenology 2011;76:409-418. doi: 10.1016/j.theriogenology.2011.02.016


71.
Hernández-Avilés C, Ramírez-Agámez L, Varner DD, et al: Effects of egg yolk level, penetrating cryoprotectant, and pre-freeze cooling rate, on the post-thaw quality of stallion sperm. Anim Reprod Sci 2023;248:107162. doi: 10.1016/j.anireprosci.2022.107162


72.
Alvarenga MA, Papa FO, Landim-Alvarenga FC, et al: Amides as cryoprotectants for freezing stallion semen: a review. Anim Reprod Sci 2005;89:105-113. doi: 10.1016/j.anireprosci.2005.07.001


73.
Melo CM, Zahn FS, Martin I, et al: Influence of semen storage and cryoprotectant on post-thaw viability and fertility of stallion spermatozoa. J Equine Vet Sci 2007;27:171–175. doi: 10.1016/j.jevs.2007.02.008


74.
Alvarenga MA, Papa FO, Ramires Neto C: Advances in stallion semen cryopreservation. Vet Clin North Am: Equine Pract 2016;32:521-530. doi: 10.1016/j.cveq.2016.08.003


75.
Graham JK: Evaluation of alternative cryoprotectants for preserving stallion spermatozoa. Proc Int Cong Anim Reprod 2000;2:307.


76.
Alvarenga MA, Graham KJ, Keith SL, et al: Alternative cryoprotectors for freezing stallion spermatozoa. Proc Int Cong Anim Reprod 2000;2:157.


77.
Medeiros ASL, Gomes GM, Carmo MT, et al: Cryopreservation of stallion sperm using different amides. Theriogenology 2002;58:273-276. doi: 10.1016/S0093-691X(002)00898-1


78.
Gomes GM, Jacob JCF, Medeiros ASL, et al: Improvement of stallion spermatozoa preservation with alternative cryoprotectants for the Mangalarga Marchador breed. Theriogenology 2002;58:277-279. doi: 10.1016/S0093-691X(02)00899-3


79.
Vidament M, Daire C, Yvon JM, et al: Motility and fertility of stallion semen frozen with glycerol and/or dimethylformamide. Theriogenology 2002;58:249-251. doi: 10.1016/S0093-691X(02)00854-3


80.
Squires EL, Keith SL, Graham JK: Evaluation of alternative cryoprotectants for preserving stallion spermatozoa. Theriogenology 2004;62:1056-1065. doi: 10.1016/j.theriogenology.2003.12.024


81.
Moore AI, Squires EL, Bruemmer JE, et al: Effect of cooling rate and cryoprotectant on the cryosurvival of equine spermatozoa. J Equine Vet Sci 2006;26:215-218. doi: 10.1016/j.jevs.2006.03.003


82.
Moffet PD, Bruemmer JE, Card C, et al: Comparison of dimethyl formamide and glycerol for cryopreservation of equine spermatozoa. Soc Theriogenol 2003;42.


83.
Morillo Rodriguez A, Balao da Silva C, Macías-García B, et al. Dimethylformamide improves the in vitro characteristics of thawed stallion spermatozoa reducing sublethal damage. Reprod Dom Anim 2012;47:995-1002. doi: 10.1111/j.1439-0531-2012.02005.x

84. Darr CR, Cortopassi GA, Datta S, et al: Mitochondrial oxygen consumption is a unique indicator of stallion spermatozoal health and varies with cryopreservation media. Theriogenology 2016;86:1382-1392. doi: 10.1016/j.theriogenology.2016.04.082


85.
Mocé E, Blanch E, Tomás C, et al: Use of cholesterol in sperm cryopreservation: present moment and perspectives to future. Reprod Dom Anim 2010;45:57-66. doi: 10.1111/j.1439-0531.2010.01635.x


86.
Combes GB, Varner DD, Schroeder F, et al: Effect of cholesterol on the motility and plasma membrane integrity of frozen equine spermatozoa after thawing. J Reprod Fertil 2000;56:127-132.


87.
Zahn FS, Papa FO, Dell’Aqua Jr, JA: Cholesterol incorporation on equine sperm membrane: effects on post-thaw sperm parameters and fertility. Theriogenology 2002;58:237-249. doi: 10.1016/S0093-691X(02)00762-8


88.
Moore AI, Squires EL, Graham JK: Adding cholesterol to the stallion sperm plasma membrane improves cryosurvival. Cryobiology 2005;51:241-249. doi: 10.1016/j.cryobiol.2005.07.004


89.
Spizziri BE, Fox MH, Bruemmer JE, et al: Cholesterol-loaded-cyclodextrins and fertility potential of stallions spermatozoa. Anim Reprod Sci 2010;118:255-264. doi: 10.1016/j.anireprosci.2009.08.001


90.
Loomis PR, Squires EL, Skaife J, et al: The relationship between cholesterol level in stallion sperm and their cryosurvival rate. J Equine Vet Sci 2018;66:44. doi: 10.1016/j.jevs.2018.05.020


91.
Blommaert D, Franck T, Donnay I, et al: Substitution of egg yolk by a cyclodextrin-cholesterol complex allows a reduction of the glycerol concentration in the freezing medium of equine sperm. Cryobiology 2016;72:27-32. doi: 10.1016/j.cryobiol.2015.11.008


92.
World Breeding Federation of Sport Horses: Semen Standards. Available from: www.wbfsh.org/files/Semen%20standards.pdf [cited 20 August 2020].


93.
Tischner M: Evaluation of deep-frozen semen in stallions. J Reprod Fertil 1979;27:53-59.


94.
Müller Z: Fertility of frozen equine semen. J Reprod Fertil 1982;32:47-51.


95.
Kenney RM, Hurtgen J, Pierson R, et al: Society for Theriogenology Manual for Clinical Fertility Evaluation of the Stallion, Society for Theriogenology, Hastings, Neb: Society for Theriogenology; 1983. 100p.


96.
Pickett BW, Back DG: Procedures for preparation, collection, evaluation, and insemination of stallion semen. Fort Collins: Colorado State University Experiment Station; 1973.


97.
Hoogewijs MK, Govaere JL, Rijsselaere T, et al: Influence of technical settings on CASA motility parameters of frozen-thawed stallion semen. Proc Am Assoc Equine Pract 2009;55:336-337.


98.
Bielanski W: Characteristics of the semen of stallions. Macro and microscopic investigations with estimation of fertility. Mem Acad Pol Sci Let B 1951;16.


99.
Nishikawa Y, Waide Y, Onuma H: Studies on artificial insemination in the horse. VI. Morphological studies on horse spermatozoa. Bull Natl Inst Agric Sci Tokyo Ser G 1951;129.


100.
Dott HM: Morphology of stallion spermatozoa. J Reprod Fertil 1975;23:41-46.


101.
Bielanski W, Kaczmarski F: Morphology of spermatozoa in semen from stallions of normal fertility. J Reprod Fertil 1979;27:39-45.


102.
Foster ML, Varner DD, Hinrichs K, et al: Agreement between measures of total motility and membrane integrity in stallion sperm. Theriogenology 2011;5:1499-1505. doi: 10.1016/j.theriogenology.2010.12.011


103.
Kiser AM, Brinsko SP, Love CC, et al: Relationship of sperm quality to fertility after 4 days of cooled storage of equine semen. J Equine Vet Sci 2014;34:602-605. doi: 10.1016/j.jevs.2013.11.007


104.
Hernández-Avilés C, Ramírez-Agámez L, Varner DD, et al: Factors affecting the analysis and interpretation of sperm quality in frozen/thawed stallion semen. Theriogenology 2024;218:35-44. doi: 10.1016/j.theriogenology.2024.01.039


105.
Merkies K, Chenier T, Plante C, et al: Assessment of stallion spermatozoa viability by flow cytometry and light microscope analysis. Theriogenology 2000;54:1215-1224. doi: 10.1016/s0093-691x(00)00428-3


106.
Hansen C, Vermeiden T, Vermeiden JPW, et al: Comparison of FACSCount AF system, Improved Neubauer hemocytometer, Corning 254 photometer, SpermVision, UltiMate, and NucleoCounter SP100 for determination of sperm concentration in boar semen. Theriogenology 2006;66:2188-2194. doi: 10.1016/j.theriogenology.2006.05.020


107.
Anzar M, Kroetsch T, Buhr MM: Comparison of different methods for assessment of sperm concentration and membrane integrity with bull semen. J Androl 2009;30:661-668. doi: 10.2164/jandrol.108.007500


108.
Foster ML, Love CC, Varner DD, et al: Comparison of methods for assessing integrity of equine sperm membranes. Theriogenology 2011;76:334-341. doi: 10.1016/j.theriogenology.2011.02.012


109.
Love CC: Relationship between sperm motility, morphology and the fertility of stallions. Theriogenology 2011;6:547-557. doi: 10.1016/j.theriogenology.2011.03.007


110.
Barrier Battut I, Kempfer A, Lemasson N, et al: Prediction of the fertility of stallion frozen-thawed semen using a combination of computer-assisted motility analysis, microscopical observation and flow cytometry. Theriogenology 2017;97:186-200. doi: 10.1016/j.theriogenology.2017.04.36


111.
Hinrichs K: Assisted reproductive techniques in mares. Reprod Dom Anim 2018;53:4-13. doi: 10.1111/rda.13259


112.
Stout TAE: Clinical application of in vitro embryo production in the horse. J Equine Vet Sci 2020;89:103011. doi: 10.1016/j.jevs.2020.103011


113.
Claes A, Stout TAE: Success rate in a clinical equine in vitro embryo production program. Theriogenology 2022;187:215-218. doi: 10.1016/j.theriogenology.2022.04.019


114.
Rader K, Choi YH, Hinrichs K: Intracytoplasmic sperm injection, embryo culture, and transfer of in vitro-produced blastocysts. Vet Clin North Am Equine Pract 2016;32:401-413. doi: 10.1016/j.cveq.2016.07.003


115.
McKinnon AO, Lacham-Kaplan O, Trounson AO: Pregnancies produced from fertile and infertile stallions by intracytoplasmic sperm injection (ICSI) of single frozen-thawed spermatozoa into in vivo matured mare oocytes. J Reprod Fertil 2000;56:513-517.


116.
Li X, Morris LHA, Allen WR: Influence of co-culture during maturation on the developmental potential of equine oocytes fertilized by intracytoplasmic sperm injection (ICSI). Reproduction 2001;12:925-932. doi: 10.1530/rep.0.1210925


117.
Choi YH, Love CC, Love LB, et al: Developmental competence in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection with fresh or frozen-thawed spermatozoa. Reproduction 2002;123:455-465. doi: 10.1530/rep.0.1230455


118.
Hinrichs K, Choi YH, Love LB, et al: Effects of holding time and media on meiotic and developmental competence of horse oocytes. Theriogenology 2002;58:675-678. doi: 10.1016/S0093-691X(02)00802-6


119.
Tremoleda JL, Colenbrander B, Stout TAE, et al: Reorganization of the cytoskeleton and chromain in horse oocytes following intracytoplasmic sperm injection. Theriogenology 2002;58:697-700. doi: 10.1016/S0093-691X(02)00794-X


120.
Galli C, Crotti G, Turini P, et al: Frozen-thawed embryos produced by Ovum Pick Up of immature oocytes and ICSI are capable to establish pregnancies in the horse. Theriogenology 2002;58:705-708. doi: 10.1016/S0093-691X(02)00771-9


121.
Lazzari G, Crotti G, Turini P, et al: Equine embryos at the compacted morula and blastocyst stage can be obtained by intracytoplasmic sperm injection (ICSI) of in vitro matured oocytes with frozen-thawed spermatozoa from semen of different fertilities. Theriogenology 2002;58:709-712. doi: 10.1016/S0093-691X(02)00777-X


122.
Campos-Chillon F, Altermatt JL: Effect of cooled vs frozen stallion semen on the efficiency of equine in vitro embryo production. J Equine Vet Sci 2023;125:104634. doi: 10.1016/j.jevs.2023.104634


123.
Choi YH, Love CC, Varner DD, et al: Equine blastocyst development after intracytoplasmic injection of sperm subjected to two freeze-thaw cycles. Theriogenology 2006;65:808-819. doi: 10.1016/j.theriogenology.2005.04.035


124.
Love CC, Kenney RM: The relationship of increased susceptibility of sperm DNA to denaturation and fertility in the stallion. Theriogenology 1998;50:955-972. doi: 10.1016/s0093-691x(98)00199-x


125.
Ramírez-Agámez L, Hernández-Avilés C, Varner DD, et al: Sperm factors associated with the production of equine blastocysts by intracytoplasmic sperm injection (ICSI) using frozen/thawed semen. Theriogenology 2023;195:85-92. doi: 10.1016/j.theriogenology.2022.10.014


126.
Palermo G, Joris H, Devroey P, et al: Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Lancet 1992;340:17-18. doi: 10.1016/0140-6736(92)92425-f


127.
Liu DY, Bourne H, Baker HWG: Fertilization and pregnancy with acrosome intact sperm by intracytoplasmic sperm injection in patients with disordered zona pellucida induced acrosome reaction. Fertil Steril 1995;64:116-121. doi: 10.1016/S0015-0228(16)57666-3


128.
Tournaye H, Liu J, Nagy Z, et al: The use of testicular sperm for intracytoplasmic sperm injection in patients with necrozoospermia. Fertil Steril 1996;66:331-334. doi: 10.1016/s0015-0282(16)58462-3


129.
Boulet SL, Mehta A, Kissin DM, et al: Trends in use of and reproductive outcomes associated with intracytoplasmic sperm injection. J Am Assoc Med 2015;313:255-263. doi: 10.1001/jama.2014.17985


130.
Practice Committees of the American Society for Reproductive Medicine and the Society for Assisted Reproductive Technology Intracytoplasmic Sperm Injection (ICSI) for non-male factor indications: A committee opinion. Fertil Steril 2020;114:239-245. doi: 10.1016/j.fertnstert.2020.05.032


131.
Dang VQ, Vuong LN, Luu TM, et al: Intracytoplasmic sperm injection versus conventional in-vitro fertilisation in couples with infertility in whom the male partner has normal total sperm count and motility: an open-label, randomised controlled trial. Lancet 2021;397:1554-1563. doi: 10.1016/S0140-6736(21)00535-3


132.
Bungum M, Humaidan P, Axmon A, et al: Sperm DNA integrity assessment in prediction of assisted reproduction technology outcome. Hum Reprod 2006;22:174-179. doi: 10.1093/humrep/del326


133.
Osman A, Alsomait H, Seshadri S, et al: The effect of sperm DNA fragmentation on live birth after IVF or ICSI: a systematic review and meta-analysis. Reprod Biomed Online 2015;30:120-127. doi: 10.1016/j.rbmo.2014.10.018


134.
Li F, Duan X, Li M, et al: Sperm DNA fragmentation index affect pregnancy outcomes and offspring safety in assisted reproductive technology. Sci Rep 2024;14:356. doi: 10.1038/s41598-023-45091-6


135.
Setti AS, Braga DAPF, Provenza RR, et al: Oocyte ability to repair sperm DNA fragmentation: the impact of maternal age on intracytoplasmic sperm injection outcomes. Fertil Steril 2021;116:123-129. doi: 10.1016/j.fertnstert.2020.10.045


136.
Gonzalez-Castro RA, Carnevale EM: Association of equine sperm population parameters with outcome of intracytoplasmic sperm injections. Theriogenology 2018;119:114-120. doi: 10.1016/j.theriogenology.2018.06.027


137.
Gibb Z, Clulow JR, Maclellan LJ, et al: Storing stallion sperm in SpermSafe™ at 17°C may improve fertility by reducing mPTP formation. J Equine Vet Sci 2023;125:104589. doi: 10.1016/j.jevs.2023.104589


138.
Consuegra C, Crespo F, Dorado J, et al: Vitrification of stallion sperm using 0.25 ml straws: effects of volume, concentration and carbohydrates (sucrose/trehalose/raffinose). Anim Reprod Sci 2019;206:69-77. doi: 10.1016/j.anireprosci.2019.05.009


139.
Diaz-Jimenez M, Rota A, Dorado J, et al: First pregnancies in jennies with vitrified donkey semen using a new warming method. Animal 2021;15:100097. doi: 10.1016/j.animal.2020.100097


140.
Felix MR, Turner RM, Dobbie T, et al: Successful in vitro fertilization in the horse: production of blastocysts and birth of foals after prolonged sperm incubation for capacitation. Biol Reprod 2022;107:1551-1564. doi: 10.1093/biolre/ioac172
Published
2024-12-13
How to Cite
Hernández-Avilés , C. (2024). Stallion sperm cryopreservation: concepts in cryopreservation-induced sperm damage, processing, analysis, and utilization in assisted reproductive technologies. Clinical Theriogenology, 16. https://doi.org/10.58292/CT.v16.10832
Section
Review Reports