Volume 13, Issue 10 (10-2015)                   IJRM 2015, 13(10): 633-644 | Back to browse issues page

XML Persian Abstract Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Ali Mohamed M S. Slow cryopreservation is not superior to vitrification in human spermatozoa; an experimental controlled study. IJRM 2015; 13 (10) :633-644
URL: http://ijrm.ir/article-1-594-en.html
Slow cryopreservation is not superior to vitrification in human spermatozoa; an experimental controlled study
Mohamed Sheata Ali Mohamed *
Women Hospital, Faculty of Medicine, University of Cologne. Kerpener Straße 62, 50937 Cologne, Germany , Mohammed.shehatta1@gmail.com
Abstract:   (2679 Views)
Background: Spermatozoa cryopreservation is used for the management of infertility and some other medical conditions. The routinely applied cryopreservation technique depends on permeating cryoprotectants, whose toxic effects have raised the attention towards permeating cryoprotectants-free vitrification technique.
Objective: To compare between the application of slow cryopreservation and vitrification on human spermatozoa.
Materials and Methods: This was an experimental controlled study involving 33 human semen samples, where each sample was divided into three equal parts; fresh control, conventional slow freezing, and permeating cryoprotectants-free vitrification. Viability and mitochondrial membrane potential (MMP) of control and post-thawing spermatozoa were assessed with the sperm viability kit and the JC-1 kit, respectively, using fluorescence-activated cell sorting analysis.
Results: Significant reduction of the progressive motility, viability and MMP was observed by the procedure of freezing and thawing, while there was not any significant difference between both cryopreservation techniques. Cryopreservation resulted in 48% reduction of the percentage of viable spermatozoa and 54.5% rise in the percentage of dead spermatozoa. In addition, high MMP was reduced by 24% and low MMP was increased by 34.75% in response to freezing and thawing. Progressive motility of spermatozoa was correlated significantly positive with high MMP and significantly negative with low MMP in control as well as post-thawing specimens (r=0.8881/ -0.8412, 0.7461/ -0.7510 and 0.7603/ -0.7839 for control, slow and vitrification respectively, p=0.0001).  
Conclusion: Although both cryopreservation techniques have similar results, vitrification is faster, easier and associated with less toxicity and costs. Thus, vitrification is recommended for the clinical application.
Keywords: Human spermatozoa, Cryopreservation, Vitrification, Sperm viability, Mitochondrial membrane potential
Full-Text [PDF 539 kb]   (697 Downloads) |   |   Full-Text (HTML)  (468 Views)  
Type of Study: Original Article |
References
1. World Health Organization. WHO laboratory manual for the examination and processing of human semen. 5th Ed. WHO Press. 2010.
2. Isachenko E, Isachenko V, Weiss JM, Kreienberg R, Katkov II, Schulz M, et al. Acrosomal status and mitochondrial activity of human spermatozoa vitrified with sucrose. Reproduction 2008; 136: 167-173. [DOI:10.1530/REP-07-0463]
3. Isachenko E, Isachenko V, Sanchez R, Katkov II, Kreienberg R. Cryopreservation of spermatozoa: Old routine and new perspectives. In: Donnez J, Kim SS. Principles and practice of fertility preservation. Cambridge, UK: Cambridge University Press, 2010; 177-199.
4. Isachenko V, Maettner R, Petrunkina AM, Sterzik K, Mallmann P, Rahimi G, et al. Cryoprotectant free vitrification of human spermatozoa in large (to 0.5 mL) volume: novel technology. Clin Lab 2011; 57: 643-650.
5. Vidament M, Vincent P, Martin FX, Magistrini M, Blesbois E. Differences in ability of Jennies and Mares to conceive with cooled and frozen semen containing glycerol or not. Anim Reprod Sci 2009; 112: 22-35. [DOI:10.1016/j.anireprosci.2008.03.016]
6. 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: 323-326. [DOI:10.1071/RD11061]
7. Isachenko EP, Mallmann G, Rahimi J. Risopatròn M, Schulz V, Isachenko R, et al. Vitrification technique- new possibilities for male gamete low-temperature storage, Current Frontiers in Cryobiology, Chapter 2. Pages 41- 76. Prof. Igor Katkov Ed. Publisher: InTech, 2012.
8. Isachenko V, Maettner R, Petrunkina AM, Sterzik K, Mallmann P, Rahimi G, et al. Vitrification of human ICSI/IVF spermatozoa without cryoprotectants: new capillary technology. J Androl 2012; 33: 462-468. [DOI:10.2164/jandrol.111.013789]
9. Yoon SJ, Kwon WS, Rahman MS, Lee JS, Pang MG. A novel approach to identifying physical markers of cryo-damage in bull spermatozoa. PLoS One 2015; 10: e0126232. [DOI:10.1371/journal.pone.0126232]
10. Pukazhenthi BS, Nagashima J, Travis AJ, Costa GM, Escobar EN, França LR, et al. Slow freezing, but not vitrification supports complete spermatogenesis in cryopreserved, neonatal sheep testicular xenografts. PLoS One 2015; 10: e0123957. [DOI:10.1371/journal.pone.0123957]
11. Jiménez-Rabadán P, García-Álvarez O, Vidal A, Maroto-Morales A, Iniesta-Cuerda M, Ramón M, et al. Effects of vitrification on ram spermatozoa using free-egg yolk extenders. Cryobiology 2015; 71: 85-90. [DOI:10.1016/j.cryobiol.2015.05.004]
12. Moraes EA, Matos WC, Graham JK, Ferrari WD Jr. Cholestanol-loaded-cyclodextrin improves the quality of stallion spermatozoa after cryopreservation. Anim Reprod Sci 2015; 158: 19-24. [DOI:10.1016/j.anireprosci.2015.04.004]
13. Agha-Rahimi A, Khalili MA, Nabi A, Ashourzadeh S. Vitrification is not superior to rapid freezing of normozoospermic spermatozoa: effects on sperm parameters, DNA fragmentation and hyaluronan binding. Reprod Biomed Online 2014; 28: 352-358. [DOI:10.1016/j.rbmo.2013.11.015]
14. Esteves SC, Sharma RK, Thomas AJ Jr, Agarwal A. Improvement in motion characteristics and acrosome status in cryopreserved human spermatozoa by swim-up processing before freezing. Hum Reprod 2000; 15: 2173-2179. [DOI:10.1093/humrep/15.10.2173]
15. Valcarce DG, Cartón-García F, Herráez MP, Robles V. Effect of cryopreservation on human sperm messenger RNAs crucial for fertilization and early embryo development. Cryobiology 2013; 67: 84- 90. [DOI:10.1016/j.cryobiol.2013.05.007]
16. Maettner R1, Sterzik K, Isachenko V, Strehler E, Rahimi G, Alabart JL, et al. Quality of human spermatozoa: relationship between high-magnification sperm morphology and DNA integrity. Andrologia 2014; 46: 547-555. [DOI:10.1111/and.12114]
17. Quill AT, Garbers LD. Sperm Motility Activation and Chemoattraction. In Daniel M. Hardy. Fertilization. Carlifornia: Academic press 2002: 29. [DOI:10.1016/B978-012311629-1/50004-8]
18. Figueroa E, Merino O, Risopatrón J, Isachenko V, Sánchez R, Effer B, et al. Effect of seminal plasma on Atlantic salmon (Salmo salar) sperm vitrification. Theriogenology 2015; 83: 238-245. [DOI:10.1016/j.theriogenology.2014.09.015]
19. Gao DY, Liu C, McGann LE, Watson PF, Kleinhans FW, Mazur P, et al. Prevention of osmotic injury to human spermatozoa during addition and removal of glycerol. Hum Reprod 1995; 10: 1109-1122. [DOI:10.1093/oxfordjournals.humrep.a136103]
20. Isachenko V, Isachenko E, Sanchez R, Dattena M, Mallmann P, Rahimi G. Cryopreservation of whole ovine ovaries with pedicles as a model for human: parameters of perfusion with simultaneous saturations by cryoprotectants. Clin Lab 2015; 61: 415-420. [DOI:10.7754/Clin.Lab.2014.140919]
21. O'Connell M, McClure N, Lewis SEM. The effect of cryopreservation on sperm morphology, motility and mitochondrial function. Hum Reprod 2002; 17: 704-709. [DOI:10.1093/humrep/17.3.704]
22. Chatterjee S, Gagnon C. Reproduction of reactive oxygen species by spermatozoa undergoing cooling, freezing, and thawing. Mol Reprod Dev 2001; 59, 451-458. [DOI:10.1002/mrd.1052]
23. Mierke CT, Kollmannsberger P, Zitterbart DP, Diez G, Koch TM, Marg S, et al. Vinculin facilitates cell invasion into three-dimensional collagen matrices. J Biol Chem 2010; 285: 13121-13130. [DOI:10.1074/jbc.M109.087171]
24. Isachenko E, Isachenko V, Katkov II, Rahimi G, Schondorf T, Mallmann P, et al. DNA integrity and motility of human spermatozoa after standard slow freezing versus cryoprotectant-free vitrification. Hum Reprod 2004; 19; 932-939. [DOI:10.1093/humrep/deh194]
25. Isachenko V, Isachenko E, Katkov II, Montag M, Dessole S, Nawroth F, et al. Cryoprotectant-free cryopreservation of human spermatozoa by vitrification and freezing in vapor: effect on motility, DNA integrity, and fertilization ability. Biol Reprod 2004; 71: 1167–1173. [DOI:10.1095/biolreprod.104.028811]
26. Kim S, Agca C, Agca Y. Changes in rat spermatozoa function after cooling, cryopreservation and centrifugation processes. Cryobiology 2010; 65: 215–223. [DOI:10.1016/j.cryobiol.2012.06.006]
27. Watson PF. The causes of reduced fertility with cryopreserved semen. Anim Reprod Sci 2000; 60–61; 481–492. [DOI:10.1016/S0378-4320(00)00099-3]
28. Pettitt MJ, Buhr MM. Extender components and surfactants affect boar sperm function and membrane behavior during cryopreservation. J Androl 1998; 19: 736-746.
29. Waberski D, Henning H, Petrunkina AM. Assessment of storage effects in liquid preserved boar semen. Reprod Domest Anim 2011; 46: 45–48. [DOI:10.1111/j.1439-0531.2011.01836.x]
30. Pe-a AI, Johannisson A, Linde-Forsberg C. Validation of flow cytometry for assessment of viability and acrosomal integrity of dog spermatozoa and for evaluation of different methods of cryopreservation. J Reprod Fertil 2001; 57: 371- 376.
31. Holt WV. Basic aspects of frozen storage semen. Anim Reprod Sci 2000; 62: 3-22. [DOI:10.1016/S0378-4320(00)00152-4]
32. Amann RP. Cryopreservation of sperm. In: Knobil E, Neill JD (eds). Encyclopedia of reproduction. Academic Press, Burlington, MA. 1999; 773–783.
33. Amaral S, Redmann K, Sanchez V, Mallidis C, Ramalho-Santos J, Schlatt S. UVB irradiation as a tool to assess ROS-induced damage in human spermatozoa. Andrology 2013; 1: 707-714. [DOI:10.1111/j.2047-2927.2013.00098.x]
34. de Lamirande E, O'Flaherty C. Sperm activation: role of reactive oxygen species and kinases. Biochim Biophys Acta 2008; 1784: 106-115. [DOI:10.1016/j.bbapap.2007.08.024]
35. Wang MJ, Ou JX, Chen GW, Wu JP, Shi HJ, O WS, et al. Does prohibitin expression regulate sperm mitochondrial membrane potential, sperm motility, and male fertility? Antioxid Redox Signal 2012; 17: 513-519. [DOI:10.1089/ars.2012.4514]
36. Varisli O, Scott H, Agca C, Agca Y. The effects of cooling rates and type of freezing extenders on cryosurvival of rat sperm. Cryobiology 2013; 67: 109-116. [DOI:10.1016/j.cryobiol.2013.05.009]
37. Yamashiro H, Eimei S. Cryopreservation of rat sperm. Current Frontiers in Cryobiology. Pages 165- 178. Prof. Igor Katkov Ed. Publisher: InTech, 2012. [DOI:10.5772/32599]
38. Ghaleno LR, Valojerdi MR, Janzamin E, Chehrazi M, Sharbatoghli M, Yazdi RS. Evaluation of conventional semen parameters, intracellular reactive oxygen species, DNA fragmentation and dysfunction of mitochondrial membrane potential after semen preparation techniques: a flow cytometric study. Arch Gynecol Obstet 2014; 289: 173-180. [DOI:10.1007/s00404-013-2946-1]
39. Kuleshova LL, MacFarlane DR, Trounson AO, Shaw JM. Sugars exert a major influence on the vitrification properties of ethylene glycol-based solutions and have a low toxicity to embryos and oocytes. Cryobiology 1999; 38: 119–130. [DOI:10.1006/cryo.1999.2153]
40. Koshimoto C, Mazur P. Effects of cooling and warming rate to and from -70°C, and effect of further cooling from -70 to -196°C on the motility of mouse spermatozoa. Biol Reprod 2002; 66: 1477–1484. [DOI:10.1095/biolreprod66.5.1477]
Send email to the article author

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Designed & Developed by : Yektaweb