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Cryosurvival and In Vitro Fertilizing Capacity Postthaw Is Improved When Boar Spermatozoa Are Frozen in the Presence of Seminal Plasma From Good Freezer Boars

JUAN J. CALVETE

LIBIA SANZ

TERESA MUIñO-BLANCO

JOSÉ A. CEBRIÁN-PÉREZ

From the ^* Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, University of Murcia, Murcia, Spain; Institute of Biomedicine, Spanish National Research Council, CSIC, Valencia, Spain; and Department of Biochemistry and Molecular and Cell Biology, Faculty of Veterinary Medicine, University of Zaragoza, Spain.

Correspondence to: Jordi Roca, Departamento de Medicina y Cirugía Animal, Facultad de Veterinaria, Universidad de Murcia, Campus de Espinardo, 30071 Murcia, Spain (e-mail: roca{at}um.es).

The study evaluated the protective effect of seminal plasma (SP) added to freezing extender against cryopreservation injuries to boar spermatozoa. Pooled sperm-rich fractions collected from 9 fertile boars were frozen in 0.5-mL straws after being extended in a conventional freezing extender either alone or supplemented with 5% of SPs (SP1–SP4) collected from the sperm-rich fractions (diluted 1:1, vol/vol, in Beltsville Thawing Solution extender) from 4 boars (1–4) with known sperm cryosurvival (poor, moderate, and good sperm freezers). Cryopreservation injuries were assessed in terms of postthaw sperm motility (assessed by computer-assisted sperm analysis), viability (plasma membrane and acrosome integrity assessed simultaneously by flow cytometry), membrane lipid peroxidation (malondialdehyde [MDA] production), and the ability of thawed spermatozoa to fertilize in vitro–matured homologous oocytes. The addition of SP from good sperm freezers (SP3 and SP4) improved (P < .01) the motility and viability of thawed spermatozoa without any influence on MDA production. Moreover, SP from good sperm freezers also increased (P < .05) the percentage of penetrated (SP3) and polyspermic oocytes (SP4) with respect to the control. Neither the total amount of SP proteins, protein profiles, nor antioxidant capacity of the different SPs were related to the various cryosurvival/fertilizing capacities of the processed spermatozoa.

     Key words: Cryopreservation, pig

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