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

  1. Elvira Matilla 1
  2. Lauro González-Fernández 2
  3. Felipe Martínez-Pastor 3
  4. Nuria Hernández 1
  5. Carolina Tobajas 1
  6. Violeta Calle-Guisado 4
  7. José Mijares 1
  8. Francisco M. Sánchez-Margallo 1
  9. Ignacio S. Álvarez 4
  10. Beatriz Macías-García 1
  1. 1 Jesús Usón Minimally Invasive Surgery Centre (CCMIJU), Cáceres, España
  2. 2 University of Porto, Portugal
  3. 3 University of León, España
  4. 4 University of Extremadura, España
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Revista:
Spanish journal of agricultural research

ISSN: 1695-971X,  2171-9292

Año de publicación: 2017

Volumen: 15

Número: 3

Tipo: Artículo

DOI: 10.5424/SJAR/2017153-11463 DIALNET GOOGLE SCHOLAR lock_openDialnet editor

Otras publicaciones en: Spanish journal of agricultural research

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.

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Información de financiación

Funding: Spanish Ministry of Economy and Competitiveness (CDTI Reprobeb Project ITC-2015-1342; Juan de la Cierva Grant IJCI-2014-19428 to BMG; Grant PTA-2012-7650 to CT); Jesús Usón Minimally Invasive Surgery Centre (Grant to EM); Agencia Estatal de Investigación (AEI; Spanish Ministry of Economy, Industry and Competitiveness) and FEDER (AGL2015-73249-JIN); Fundação para a Ciência e a Tecnologia (Portugal)/ESF/Portuguese Ministry for Science, Technology and Higher Education (Grant SFRH/BPD/85532/2012 to LGF); Spanish Ministry of Education, Culture and Sport (Grant FPU-014/03449 to VCG). Competing interests: The authors declare no conflicts of interest. Correspondence should be addressed to Beatriz Macías-García: bmacias@ccmijesususon.com

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Fuente de los datos: Dialnet