Long-term preservation of freeze-dried rabbit sperm by adding rosmarinic acid and different chelating agents

Domingo, P. ; Olaciregui, M. ; Gonzalez, N. (Universidad de Zaragoza) ; De Blas, I. (Universidad de Zaragoza) ; Gil, L. (Universidad de Zaragoza)

Resumen: Freeze-drying (FD) technique has been applied as an alternative technology to preserve gene resources to allow simple sperm preservation and shipment at 4 degrees C. Nevertheless, DNA sperm might be damaged by mechanical or oxidative stress throughout FD procedure. Therefore, suitable protection to maintain DNA integrity is required. The aim of this study was to determine the effect of rosmarinic acid (RA) as an antioxidant and two chelating agents (EGTA and EDTA) on the DNA integrity of freeze-dried rabbit sperm after storage of the samples at 4 degrees C and room temperature for 8 months. Rabbit sperm were freeze-dried in basic medium (10 mM Tris-HCl buffer and 50 mM NaCl) supplemented with 50 mM EGTA (1), 50 mM EGTA plus 105 mu M RA (2), 50 mM EDTA (3) or 50 mM EDTA plus 105 mu M RA (4). Semen samples were kept at 4 degrees C and room temperature during 8 months. After rehydration, DNA integrity was evaluated with Sperm Chromatin Dispersion test observing that DNA fragmentation was higher when semen samples were freeze-dried with EGTA (10.9%) than with EDTA (4.1%) (p < 0.01). Furthermore, RA acted better under adverse conditions and no significant differences were found in temperature storage. Summarizing, FD is a method that can allow simple gene resources preservation among 4 degrees C to 25 degrees C during 8 months and transportation without the need for liquid nitrogen or dry ice. EDTA chelating agent is the most suitable media for freeze-dried rabbit sperm and the addition of RA protects the DNA against the oxidative stress caused during FD procedure.

Idioma: Inglés

DOI: 10.1016/j.cryobiol.2018.01.004

Año: 2018

Publicado en: Cryobiology 81 (2018), 174-177
ISSN: 0011-2240

Factor impacto JCR: 2.141 (2018)

Categ. JCR: BIOLOGY rank: 29 / 87 = 0.333 (2018) - Q2 - T2
Categ. JCR: PHYSIOLOGY rank: 51 / 81 = 0.63 (2018) - Q3 - T2

Factor impacto SCIMAGO: 0.711 - Agricultural and Biological Sciences (miscellaneous) (Q1) - Medicine (miscellaneous) (Q1) - Biochemistry, Genetics and Molecular Biology (miscellaneous) (Q1)

Financiación: info:eu-repo/grantAgreement/ES/DGA/FSE

Tipo y forma: Article (PostPrint)

Área (Departamento): Área Sanidad Animal (Dpto. Patología Animal)
Área (Departamento): Área Medicina y Cirugía Animal (Dpto. Patología Animal)