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    Título
    Effect of Single Layer Centrifugation Porcicoll (70%, 80% and 90%) or supplementation with reduced glutathione, seminal plasma and bovine serum albumin on frozen-thawed boar sperm
    Autor
    Crespo Félez, I.
    Castañeda Sampedro, Ana
    Sánchez, Diana I.
    Fernández Alegre, Estela
    Álvarez Rodríguez, Manuel
    Domínguez Fernández de Tejerina, Juan CarlosAutoridad BuleriaORCID
    Morrell, Jane Margaret
    Martínez Pastor, FelipeAutoridad BuleriaORCID
    Facultad/Centro
    Facultad de Ciencias Biologicas y Ambientales
    Área de conocimiento
    Biologia Celular
    Datos de la obra
    Animal Reproduction Science, 2017, vol. 187
    Editor
    Elsevier
    Fecha
    2017-12
    Abstract
    Selecting the optimal sperm population is essential for success with reproductive techniques. Porcicoll (formerly Androcoll-P) is a colloid formulation for selection of high-quality boar spermatozoa by single layer centrifugation (SLC). To date, most studies have been carried out with fresh semen and large volumes. We carried out 2 experiments to test the use of Porcicoll for thawed boar semen in small volumes. In Experiment 1, cryopreserved semen doses were thawed, split in 200-μL aliquots and layered on 1 mL of Porcicoll 70%, 80% or 90%, or buffer without colloid. We assessed sperm recovery (the proportion of the loading dose that appeared in the pellet, %), and the physiology of the selected spermatozoa (flow cytometry: Viability, apoptotic changes, capacitation, mitochondrial activity, intracellular reactive oxygen species). The most suitable proportion was Porcicoll 80%, allowing acceptable sperm recovery (16.9 ± 4.2%, compared to 70% (35.4% ± 3.0, p < 0.001) and 90% (8.2% ± 3.0, P = 0.001), and improved quality (mitochondrial activity: Porcicoll 80%: 77.7 ± 1% vs Control: 60.3 ± 0.7%, P < 0.05). In Experiment 2, we compared 3 supplements to Porcicoll 80%: 500 mM reduced glutathione (GSH), 20% seminal plasma (SP) and 0.5% bovine serum albumin (BSA). Supplementation with GSH or BSA did not cause relevant changes relative to Control. In contrast, SP induced membrane and acrosomal changes resembling capacitation, which might preclude its use in some applications, and decreased recovery (5.5% ± 1.9 vs. 24.3% ± 1.2 Control; P < 0.001). However, it could be useful prior to other applications such as in vitro fertilisation. Overall, Porcicoll is an effective colloid for isolating a high-quality population from thawed boar sperm, 80% being a balanced option for good recovery and high quality. Supplements could be useful depending on the proposed use of the spermatozoa.
    Materia
    Veterinaria
    Palabras clave
    Bovine serum albumin
    Pig
    Semen
    Seminal plasma
    Reduced glutathione
    Single-layer centrifugation
    Peer review
    SI
    URI
    http://hdl.handle.net/10612/10703
    Versión del editor
    https://www.sciencedirect.com/science/article/pii/S0378432017307327#!
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