Porcine sperm vitrification II: Spheres method
Corresponding Author
C. C. Arraztoa
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Correspondence
Claudia Cecilia Arraztoa, Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.
Email: [email protected]
Search for more papers by this authorM. H. Miragaya
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Search for more papers by this authorM. G. Chaves
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Search for more papers by this authorV. L. Trasorras
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Search for more papers by this authorM. C. Gambarotta
Cátedra de Estadística, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Search for more papers by this authorD. M. Neild
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Search for more papers by this authorCorresponding Author
C. C. Arraztoa
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Correspondence
Claudia Cecilia Arraztoa, Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina.
Email: [email protected]
Search for more papers by this authorM. H. Miragaya
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Search for more papers by this authorM. G. Chaves
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Search for more papers by this authorV. L. Trasorras
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Search for more papers by this authorM. C. Gambarotta
Cátedra de Estadística, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Search for more papers by this authorD. M. Neild
Cátedra de Teriogenología, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Instituto de Investigación y Tecnología en Reproducción Animal (INITRA), Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Buenos Aires, Argentina
Search for more papers by this authorSummary
Owing to current problems in boar sperm cryopreservation, this study proposes to evaluate vitrification in spheres as an alternative cryopreservation procedure, comparing the use or not of permeable cryoprotectants and two warming methods. Extended (n = 3; r = 4) and raw (n = 5; r = 2) porcine spermatozoa were diluted in media, in the absence or presence of either 4% dimethylformamide or 4% glycerol, to a final concentration of 5 × 106 spermatozoa/ml and vitrified using the spheres method. Two warming procedures were evaluated: a rapid method (30 s at 37°C) and an ultrarapid method (7 s at 75°C, followed by 30 s at 37°C). Percentages of total motility (phase contrast), membrane function (hypo-osmotic swelling test), acrosome integrity (phase contrast), sperm viability (6-carboxyfluorescein diacetate and propidium iodide stain), chromatin condensation (toluidine blue stain) and chromatin susceptibility to acid denaturation (acridine orange stain) were evaluated in the samples before and after vitrification. Results, analysed using Friedman's test, suggest that rapid warming of raw porcine spermatozoa vitrified without permeable cryoprotectants may preserve DNA condensation and integrity better than the other processing methods studied in this work. Hence, porcine sperm vitrification using spheres could be used to produce embryos with ICSI to further validate this method.
References
- Arav, A., Yavin, S., Zeron, Y., Natan, D., Dekel, I., & Gacitua, H. (2002). New trends in gamete's cryopreservation. Molecular and Cellular Endocrinology, 187, 77–81.
- Bianchi, I., Calderam, K., Maschio, E., Madeira, E., Ulguim, R., Corcini, C., … Correa, M. (2008). Evaluation of amides and centrifugation temperature in boar semen cryopreservation. Theriogenology, 69, 632–638.
- Binh, N. T., Van Thuan, N., & Miyake, M. (2009). Effects of liquid preservation of sperm on their ability to activate oocytes and initiate preimplantational development after intracytoplasmic sperm injection in the pig. Theriogenology, 71, 1440–1450.
- Boe-Hansen, G., Ersboll, A., Greve, T., & Christensen, P. (2005). Increasing storage time of extended boar semen reduces sperm DNA integrity. Theriogenology, 63, 2006–2019.
- Cochran, J. D., Amann, R. P., Froman, D. P., & Pickett, B. W. (1984). Effects of centrifugation, glycerol level, cooling to 5°C, freezing rate and thawing rate on the post-thaw motility of equine sperm. Theriogenology, 22, 25–38.
- Cristanelli, M., Squires, E., Amann, R., & Pickett, B. (1984). Fertility of stallion semen processed, frozen and thawed by a new procedure. Theriogenology, 22, 39–45.
- Gao, D., & Critser, J. (2000). Mechanisms of cryoinjury in living cells. ILAR Journal, 41, 187–196.
- Gao, D., Mazur, P., & Critser, J. (1997). Fundamental cryobiology of mammalian spermatozoa. In A. M. Karow, & J. K. Critser (Eds.), Reproductive tissue banking: Scientific principles (pp. 263–328). London, UK: Academic Press.
10.1016/B978-012399770-8/50007-1 Google Scholar
- García Rosello, E., García Mengual, E., Coy, P., Alfonso, J., & Silvestre, M. (2008). Intracytoplasmic sperm injection in livestock species: An update. Reproduction in Domestic Animals, 44, 143–151.
- García Vázquez, F., García- Roselló, E., Gutiérrez-Adán, A., & Gadea, J. (2009). Effect of sperm treatment on efficiency of EGFP-expressing porcine embryos produced by ICSI-SMGT. Theriogenology, 72, 506–518.
- González, L., Campi, S., Cisale, H., Torres, P., García, C., & Ferrari, M. (2008). Descondensación in vitro de núcleos espermáticos porcinos. InVet, 10, 174.
- Grossfeld, R., Sieg, B., Struckmann, C., Frenzel, A., Maxwell, W. M. C., & Rath, D. (2008). New aspects of boar semen freezing strategies. Theriogenology, 70, 1225–1233.
- Harrison, R. A., & Vickers, S. E. (1990). Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa. Journal of Reproduction and Fertility, 88, 343–352.
- Isachenko, E., Isachenko, V., Katkov, I., Dessole, S., & Nawroth, F. (2003). Vitrification of mammalian spermatozoa in the absence of cryoprotectants: From past practical difficulties to present success. Reproductive Biomedicine Online, 6, 191–200.
- Isachenko, V., Isachenko, E., Katkov, I. I., Montag, M., Dessole, S., Nawroth, F., & van der Ven, H. (2004). Cryoprotectant-free cryopreservation of human spermatozoa by vitrification and freezing in vapor: Effect on motility, DNA integrity, and fertilization ability. Biology of Reproduction, 71, 1167–1173.
- Isachenko, V., Isachenko, E., Petrunkina, A. M., & Sanchez, R. (2012). Human spermatozoa vitrified in the absence of permeable cryoprotectants: Birth of two healthy babies. Reproduction, Fertility, and Development, 24, 323–326.
- Isachenko, E., Isachenko, V., Weiss, J., Kreienberg, R., Katkov, I., Schulz, M., … Sánchez, R. (2008). Acrosomal status and mitochondrial activity of human spermatozoa vitrified with sucrose. Reproduction, 136, 167–173.
- Jiménez-Rabadán, P., García-Álvarez, O., Vidal, A., Maroto-Morales, A., Iniesta-Cuerda, M., Ramón, M., … Soler, A. J. (2015). Effects of vitrification on ram spermatozoa using free-egg yolk extenders. Cryobiology, 71, 85–90.
- Johnson, I., Weitze, K., Fiser, P., & Maxwell, W. (2000). Storage of boar semen. Animal Reproduction Science, 62, 143–172.
- Katkov, I. (2012). Kinetic vitrification of spermatozoa of vertebrates: What can we learn from nature? In I. Katkov (Ed.), Current frontiers in cryobiology (pp. 3–40). Croatia: InTech.
10.5772/34784 Google Scholar
- Katkov, I., Isachenko, V., Isachenko, E., Kim, M., Lulatd, A., Mackaye, A., & Levine, F. (2006). Low-and high-temperature vitrification as a new approach to biostabilization of reproductive and progenitor cells. International Journal of Refrigeration, 29, 346–357.
- King, G., & Macpherson, J. (1973). A comparison of two methods for boar semen collection. Journal of Animal Science, 36, 563–565.
- Kumar Gupta, M., Jun Uhm, S., & Taek Lee, H. (2007). Cryopreservation of immature and in vitro matured porcine oocytes by solid surface vitrification. Theriogenology, 67, 238–248.
- Larsson, K., & Einarsson, S. (1976). Fertility of deep frozen boar spermatozoa: Influence of thawing diluents. Acta Veterinaria Scandinavica, 17, 43–62.
- Luyet, B., & Hodapp, R. (1938). Revival of frog spermatozoa vitrified in liquid air. Proceedings of Meeting of the Society for Experimental Biology, 39, 433–434.
- Malo, C., Gil, L., Cano, R., Martínez, F., García, A., & Jerez, R. (2012). Dimethylformamide is not better than glycerol for cryopreservation of boar semen. Andrologia, 44, 605–610.
- Mazur, P. (1984). Freezing of living cells: Mechanisms and implications. American Journal of Physiology, 247, C125–C142.
- Merino, O., Risopatrón, J., Sánchez, R., Isachenko, E., Figueroa, E., Valdebenito, I., & Isachenko, V. (2011). Fish (Oncorhynchus mykiss) spermatozoa cryoprotectant – free vitrification: Stability of mitochondrion as criterion of effectiveness. Animal Reproduction Science, 124, 125–131.
- Morris, S., & Shertzer, H. (1985). Rapid analysis of DNA strand breaks in soft tissues. Environmental and Molecular Mutagenesis, 13, 151–163.
- Nauk, V. (1991). Structure and function of spermatozoa from farm animals by cryopreservation. Shtiinca Kishinev, pp 1–198.
- Nawroth, F., Isachenko, V., Dessole, S., Rahimi, G., Farina, M., Vargiu, N., … Isachenko, E. (2002). Vitrification of human spermatozoa without cryoprotectants. CryoLetters, 23, 93–102.
- O'Conell, M., McClure, N., & Lewis, E. M. (2002). The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Human Reproduction, 17, 704–709.
- Parks, J. (1997). Hypothermia and mammalian gametes. In A. M. Karow, & J. K. Critser (Eds.), Reproductive tissue banking: Scientific principles (pp. 229–261). San Diego, CA: Academic Press.
- Parks, J., & Lynch, D. (1992). Lipid composition and thermotropic phase behavior of boar, bull, stallion and rooster sperm membrane. Cryobiology, 29, 255–266.
- Polge, C., Salamos, S., & Wilmut, I. (1970). Fertilizing capacity of frozen semen following surgical insemination. Veterinary Record, 87, 424–428.
- Pursel, V., & Johnson, L. (1974). Glutaraldehyde fixation of boar spermatozoa for acrosome evaluation. Theriogenology, 1, 63–68.
- Roca, J., Parrilla, I., Bolarin, A., Martinez, E. A., & Rodriguez-Martinez, H. (2016). Will AI in pigs become more efficient? Theriogenology, 86, 187–193.
- Rosato, M., & Iaffaldano, N. (2013). 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. Theriogenology, 79, 508–516.
- Sánchez, R., Risopatrón, J., Schulz, M., Villegas, J., Isachenko, V., Kreinberg, R., & Isachenko, E. (2011). Canine sperm vitrification with sucrose: Effect on sperm function. Andrologia, 43, 233–241.
- Swain, J. E., & Smith, G. D. (2010). Cryoprotectants. In R. C. Chian, & P. Quinn (Eds.), Fertility cryopreservation (pp. 24–38). New York, NY: Cambridge University Press.
10.1017/CBO9780511730207.005 Google Scholar
- Tejada, R., Mitchell, J., Norman, A., Marik, J., & Friedman, S. (1984). A test for the practical evaluation of male fertility by acridine orange fluorescence. Fertility and Sterility, 42, 87–91.
- Vázquez, J., Martinez, E., Martinez, P., García-Artiga, C., & Roca, J. (1997). Hypoosmotic swelling of boar spermatozoa compared to other methods for analyzing the sperm membrane. Theriogenology, 47, 913–922.
- Watson, P. (2000). The causes of reduced fertility with cryopreserved semen. Animal Reproduction Science, 60, 481–492.
- Yanagimachi, R. (2005). Male gamete contribution to the embryo. Annals of the New York Academy of Sciences, 1061, 203–207.
- Yeste, M. (2015). Recent advances in boar sperm cryopreservation: State of the art and current perspectives. Review article. Reproduction in Domestic Animals, 50, 71–79.
- Yoshida, M. (2000). Conservation of sperms: Current status and new trends. Animal Reproduction Science, 60–61, 349–355.
