Teke Spermasının Vitrifikasyonu

Year 2023, Volume: 16 Issue: 4, 521 - 529, 31.12.2023

Çiğdem Çebi Ricardo Faundez

https://doi.org/10.30607/kvj.1363108

Abstract

Vitrifikasyon metodu, soğutma hızının çok yüksek olduğu, yüksek konsatrasyonlu permeable kriyopektanların kullanıldığı bir dondurma yöntemidir. Embriyo dondurulmasında oldukça geniş uygulama alanı bulan vitrifikasyon methodunun teke spermasının saklanmasında uygulanarak sperma kalitesi üzerine vitrifikasyon methodunun etkisi belirlenmiştir. Bu amaçla 4 sağlıklı tekeden suni vajen yoluyla alınan sperma örnekleri 4 eşit gruba ayrılmıştır. Grup 1’deki sperma örnekleri %20 yumurta sarısı ve %5 gliserol içeren Tris-Sitrat modifiye solüsyonu ile konvansiyonel methodla dondurulurken, geri kalan sperm örnekleri ise 0.1 M (Grup 2), 0.25 M (Grup 3) ve 0.5 M (Grup 4) sükroz içeren Human Tubal Fluid sulandırıcısı ile sulandırılıp vitrifikasyon işlemine tabi tutulmuştur. Tüm sperma örneklerinde konvansiyonel dondurma ve vitrifikasyon işlemi sonrası standart sperma analizi (spermatozoa motilitesi, plasma membran bütünlüğü, sperm morfolojisi, kromatin bütünlüğü) yapılmıştır. Sonuç olarak, non-permeable kriyoprotektanlar kullanılarak teke spermatozoasını vitrifiye etmek için girişilen deneyimler çok düşük canlılıkla sonuçlanmıştır. Vitrifikasyon sırasında oluşan fiziksel ve kimyasal değişiklikler, spermatozoanın motilitesini, canlılığını, sperm morfolojisini olumsuz şekilde etkilemiştir.

Keywords

Sperma, Sükroz, Teke, Vitrifikasyon

Ethical Statement

Alınmıştır.

Supporting Institution

Harran Üniversitesi

Project Number

1609

Thanks

HÜBAP

Goat Sperm Vitrification

Abstract

The vitrification method is a freezing method in which the cooling rate is very high and high concentration permeable cryoprotectants are used. The vitrification method, which has a wide application area in embryo freezing, was applied to the storage of goat semen and the effect of the vitrification method on semen quality was determined. For this purpose, semen samples taken from 4 healthy goats via artificial vagina were divided into 4 equal groups. While the semen samples in Group 1 were frozen with the conventional method with Tris-sitrate modified solution containing 20% egg yolk and 5% glycerol, the remaining sperm samples were frozen with 0.1 M (Group II), 0.25 M (Group III) and 0.5 M (Group IV) was diluted with Human Tubal Fluid diluent containing sucrose and vitrification method was applied. Standard semen analysis (spermatozoa motility, plasma membrane integrity, sperm morphology, chromatin integrity) was performed on all semen samples after conventional freezing and vitrification. As a result, attempts to vitrify goat spermatozoa using non-permeable cryoprotectants have resulted in very low viability. The physical and chemical changes occurring during vitrification negatively affected the motility, viability and sperm morphology of spermatozoa.

Keywords

Goat, Semen, Sucrose, Vitrification

Project Number

1609

References

• Agha-Rahimi, A., Khalili, M.A., Nabi, A., & Ashourzadeh S. (2014). Vitrification is not superior to rapid freezing of normozoospermic spermatozoa: effects on sperm parameters, DNA fragmentation and hyaluronan binding. Reprod. Biomed, 26, 352-358.
• Antonov, A., & Ivanova, B. (2023). Canine sperm vitrification with nonpermeable cryoprotectants and coconut water extender. Anim Reprod, 23, 20(2), e20230004. doi: 10.1590/1984-3143-AR2023-0004.
• Arandoa, A., Gonzalezb, A., Delgadoa, J.V., Arrebolac, F.A., & Perez-Marín, C.C. (2017). Storage temperature and sucrose concentrations affect ram sperm quality after vitrification. Animal Reproduction Science, 181, 175-185. doi: 10.1016/j.anireprosci.2017.04.008.
• Arraztoa, C.C., Miragaya, M.H., Chaves, M.G., Trasorras, V.L., Gambarotta, M.C., & Neild, D.M. (2017). Porcine sperm vitrification II: Spheres method. Andrologia, 49(8), 1-8. https://doi: 10.1111/and.12738.
• Barbosa, B.B., Evangelista, I.T.A., Soares, A.R.B., Leão, D.L., Pereira, R.J.G., & Domingues, S.F.S. (2023). Kinetic vitrification: concepts and perspectives in animal sperm cryopreservation. Anim Reprod, 22, 20(2):e20220096. doi: 10.1590/1984-3143.
• Caturla-Sanchez, E., Sanchez-Calabuig, M., Perez-Gutierrez, J., Cerdeira, J., Castano, C., & Santiago-Moreno, J. (2018). Vitrification of dog spermatozoa: Effects of two cryoprotectants (sucrose or trehalose) and two warming procedures. Cryobiology, 80, 126. DOI: 10.1016/j.cryobiol.2017.11.001.
• Cebi Sen, C., Yumusak, N., Atilgan, H.I., Sadic, M., Koca, G., & Korkmaz, M. (2018). The protective effect of melatonin on sperm quality in rat after radioiodine treatment. Andrologia, 50, 1-7. https://doi: 10.1111/and.12962.
• Fahy, G.M., & Rall, W.F. (2007). Vitrification: an overview. In Tucker MJ, Liebermann J, eds. Vitrification in Assisted Reproduction. A User’s Manual and Troublr-Shooting Guide. London: Informa Healthcare, 1-20.
• Gao, D.Y, Ashworth, E., Watson, S.P.F., Kleinhans, F.W., Mazur, P., & Critser, J.K. (1993). Hyperosmotic tolerance of human spermatozoa: separate effects of glycerol, sodium chloride, and sucrose on spermolysis. Biol Reprod, 49, 112-123.
• Gharajelar, S.N., Sadrkhanloo, R.A., Onsori, M., & Saberivand, A. (2016). A comparative study on the effects of different cryoprotectants on the quality of canine sperm during vitrification process. Vet Res Forum, 7(3), 235-239.
• Isachenko, V., Isachenko, E., Katkov, I.I., Montag, M., Dessole, S., Nawroth, F., & Van DerVen, H. (2004a). Cryoprotectant-free cryopreservation of human spermatozoa by vitrification and freezing in vapor: effect on motility DNA integrity, and fertilization ability. Biol Reprod, 71, 1167-1173.
• Isachenko, V., Isachenko, E., Katkov, I.I., Rahimi, G., Schondorf, T., Mallmann, P., Dessole, S., & Nawroth, F. (2004b). DNA integrity and motility of human spermatozoa after standard slow freezing versus cryoprotectant-free vitrification. Hum Reprod, 19, 932-939.
• Isachenko, E., Isachenko, V., Weis, J.M., Kreienberg, R., Katkov, I.I., Schulz, M., Lulat, A.G.M.I., Risopatróm, M.J., & Sánchez, R. (2008). Acrosomal status and mitochondrial activity of human spermatozoa vitrified with sucrose. Soc Reprod Fertil, 136, 167-173.
• Isachenko, V., Maettner, R., Petrunkina, A.M., Sterzik, K., Mallmann, P., Rahimi, G., Sanchez, R., Risopatron, J., Damjanoski, I., & Isachenko, E. (2012). Vitrification of Human ICSI/IVF Spermatozoa Without Cryoprotectants: New Capillary Technology. Journal of Andrology, 33, 462-468.
• Jiménez-Rabadán, P., García-Álvarez, O., Vidal, A., Maroto-Morales, A., Iniesta-Cuerda, M., Ramón, M., Del Olmo, E., Fernández-Santos, R., Garde, J.J., & Soler, AJ. (2015). Effects of vitrification on ram spermatozoa using free-egg yolk extenders. Cryobiology, 71, 85-90.
• Kervancıoğlu, G. (2018). Kanser Hastalarında Fertilitenin Korunmasında Kullanılan Kriyoprezervasyon Yöntemlerİ. Tıp Fakültesi Klinikleri, 1(2), 17-26.
• Koshimoto, C.H., & Mazur, P. (2002). 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, 66, 1477-1484.
• Kulaksiz, R., & Daşkın, A. (2009). Farklı antioksidanlarla dondurulan Saanen teke spermasının in vitro ve in vitro değerlendirilmesi. Ankara Üniv Vet Fak Derg, 56, 201-205.
• Le, M.T., Nguyen, T.T.T., Nguyen, T.T., Nguyen, V.T., Nguyen, T.T.A., Nguyen, V.Q.H., & Cao, N.T. (2019). Cryopreservation of human spermatozoa by vitrification versus conventional rapid freezing: Effects on motility, viability, morphology and cellular defects. Eur J Obstet Gynecol Reprod Biol, 234, 14–20. doi:10.1016/j. ejogrb.2019.01.001
• Liu, J., Tanrikut, C., Wright, D.L., Lee, G.Y., Toner, M., Biggers, J.D., & Toth, T.L. (2016). Cryopreservation of human spermatozoa with minimal non-permeable cryoprotectant. Cryobiology, 73(2), 162-170.
• Lv, C., Wu, G., Hong, Q., Quan, G. (2019). Spermatozoa Cryopreservation: State of Art and Future in Small Ruminants. Biopreservation And Biobanking, 17(2), 171-182.
• Maxwell, W.M.C., Welch, G.R., & Johnson, L.A. (1997). Viability and membrane integrity of spermatozoa after dilution and flow cytometric sorting in the presence or absence of seminal plasma. Reprod Fertil Dev, 8, 1165-1178.
• McClean, R., Holt, WV., Zee, Y.P., Lisle, A., & Johnston, S.D. (2008). The effect of cryoprotectant on kangaroo sperm ultrastructure and mitochondrial function. Cryobiology, 57, 297-303.
• 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, Anim. Reprod. Sci, 124, 125-131.
• Mukaida, T., Wada, S., Takahashi, K., Pedro, P.B., An, T.Z., & Kasai, M. (1998). Vitrification of human embryos based on the assessment of suitable conditions for 8-cell embryos. Hum Reprod, 2874-2879. Nawroth, F., Isachenko, V., Dessole, S., Rahimi, G., Farina, M., Vargiu, N., Mallmann, P., Dattena, M., Capobianco, G., Peters, D., Orth, I., & Isachenko, E. (2002). Vitrification of human spermatozoa without cryoprotectants. Cryo Lett, 23, 93-102.
• Olamitibo, D.J., Dayo, O.O., Oladimeji, A.M., Mathew, A., Olajide, O., Emmanuel, O.O., Oluwafemi, A.E., Amidu, S.T., & Ayobami, I.O. (2016). Effects of avocado seed extract in different trisextenders on sperm and oxidative stress indices of vitrified goat spermatozoa. Journal of agricultural sciences, 61(4), 359-374.
• Ömür, A.D., & Çoyan, K. (2014). Sezon İçi ve Sezon Dışında Koç Spermasının Dondurulmasında Antioksidanların Etkisi. Erciyes Üniv Vet Fak Derg, 11(1), 37-42.
• Peirouvi, T., Farjah, G., Rad, J.S., & Novin, M.G. (2007). Vitrification induced apoptosis in spermatozoa from fertile and subfertile men. Iranian Journal of Reproductive Medicine, 5(3),117-120.
• Pradiee J., Sanchez-Calabuig M.J., Castano C., O'Brien E., Esteso M.C., Beltran-Brena P., Maillo V., Santiago-Moreno J., & Rizos D. (2017). Fertilizing capacity of vitrified epididymal sperm from Iberian ibex (Capra pyrenaica). Theriogenology, 108(2018), 314-320.
• Rosato M.P., & 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 freezing and vitrification. Theriogenology, 73, 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.
• Satirapod, C., Treetampinich, C., Weerakiet, S., Wongkularb, A., Rattanasiri, S., & Choktanasiri, W. (2012). Comparison of cryopreserved human sperm from solid surface vitrification and standard vapor freezing method: On motility, morphology, vitality and DNA integrity. Andrologia, 44, 786-790. doi: 10.1111/j.1439-0272.2011.01267.x
• Schulz, M., Munoz, M., Risopatron, J., & Sanchez, R. (2006): Cryopreservation of human spermatozoa by vitrification. International Journal of Morphology, 24-31.
• Shah D, Rasappan, Shila, & Karthik G. (2019). A simple method of human sperm vitrification. MethodsX, 6, 2198-2204.
• Tamburrino, L., Traini, G., Marcellini, A., Vignozzi, L., Baldi, E., & Marchiani, S. (2023). Cryopreservation of Human Cryopreservation of Human Spermatozoa: Functional, Molecular and Clinical Aspects. Int J Mol Sci, 28, 24(5), 4656. doi: 10.3390/ijms24054656.
• Tunalı G. (2014). Sperm kriyoprezervasyon teknikleri ve fertilizasyon başarısındaki rolü. Androloji Bülteni, 57, 123-128.
• Van der Horst, G., & Maree, L. (2010). SpermBlue®: A new universal stain for human and animal sperm which is also amenable to automated sperm morphology analysis, Biotechnic & Histochemistry, 84(6), 299-308.
• Vajta, G., & Nagy, Z.P. (2006). Are programmable freezers still needed in the embryo laboratory? Review on vitrification. Reprod Biomed Online, 779-796.
• Wetzels, A.M.M., Bras, M., Lens, J.W., Piederiet, M.H., Rijnders, P.M., Zeilmaker, G.H. (1996). Laboratory aspects of in vitro fertilization. Cryopreservation/Theory, 229-244.
• Zakošek, P.M,. Casal, M.L., Šterbenc, N., Virant, K.I., & Mrkun, J. (2020). Vitrification Using Soy Lecithin and Sucrose: A New Way to Store the Sperm for the Preservation of Canine Reproductive Function Animals, 10(4), 653. doi.org/10.3390/ani10040653