Outlining adequate protocols for Lidia bull epididymal storage and sperm cryopreservation:: use of glycerol, dimethylformamide and N-acetylcysteine

• Elvira Matilla ^[4] ; Lauro González-Fernández ^[1] ; Felipe Martínez-Pastor ^[2] ; Nuria Hernández ^[4] ; Carolina Tobajas ^[4] ; Violeta Calle-Guisado ^[3] ; José Mijares ^[4] ; Francisco M. Sánchez-Margallo ^[4] ; Ignacio S. Álvarez ^[3] ; Beatriz Macías-García ^[4]
  1. [1] Universidade Do Porto

     Universidade Do Porto

     Santo Ildefonso, Portugal

     
  2. [2] Universidad de León

     Universidad de León

     León, España

     
  3. [3] Universidad de Extremadura

     Universidad de Extremadura

     Badajoz, España

     
  4. [4] Jesús Usón Minimally Invasive Surgery Centre (CCMIJU), Cáceres, España

  Mostrar afiliaciones +
• Localización: Spanish journal of agricultural research, ISSN-e 2171-9292, ISSN 1695-971X, Vol. 15, Nº. 3, 2017
• Idioma: inglés
• Enlaces
  • Texto completo (pdf)
• Resumen

  • The Lidia bovine breed is an important hallmark of the Spanish cattle industry. Bulls are selected based upon aggressiveness and epididymal sperm cryopreservation is the way to obtain and store their genetics. There are not specifically designed protocols yet to perform Lidia bull sperm cryopreservation. The present study aimed to determine if a tris-fructose-citrate-egg yolk (20% v/v; TFY) extender supplemented with 7% glycerol (TFY1) or 3.5% glycerol plus 3.5% dimethylformamide (DMF; TFY2) are suitable media for cryopreservation of epididymal Lidia bull sperm. Moreover, the effect of N-acetylcysteine (NAC), a potent antioxidant, was evaluated. The epididymis were stored at 4°C for 24, 48, 72 or 96 h, and both freezing media were tested as such or supplemented with 1 or 2.5 mM of NAC. Our data demonstrated that post-thaw viability was well maintained (TFY1: 50.8% ± 1.9 at 24 h and 52.4% ± 0.8 at 96 h and TFY2: 52.6% ± 1.6 at 24 h and 56.1% ± 1.8 at 96 h; mean % ± SEM; p>0.05) as also were total and progressive sperm motility, high mitochondrial membrane potential, ROS production, DNA status and acrosomal intactness of Lidia bull sperm up to 96 h of epididymal storage, all extender variations being similar (p>0.05). In conclusion, the use of TFY medium supplemented either with 7% glycerol alone or the combination of 3.5% glycerol and 3.5% DMF were equally safe choices for epididymal Lidia bull sperm cryopreservation, and NAC addition did not significantly improve sperm post-thaw quality.