RT info:eu-repo/semantics/article
T1 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
A1 Crespo Félez, I.
A1 Castañeda Sampedro, Ana
A1 Sánchez, Diana I.
A1 Fernández Alegre, Estela
A1 Álvarez Rodríguez, Manuel
A1 Domínguez Fernández de Tejerina, Juan Carlos
A1 Morrell, Jane Margaret
A1 Martínez Pastor, Felipe
A2 Biologia Celular
K1 Veterinaria
K1 Bovine serum albumin
K1 Pig
K1 Semen
K1 Seminal plasma
K1 Reduced glutathione
K1 Single-layer centrifugation
AB 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.
PB Elsevier
YR 2019
FD 2019-05-13
LK http://hdl.handle.net/10612/10703
UL http://hdl.handle.net/10612/10703
NO Animal Reproduction Science, 2017,  vol. 187
NO P. 167-173
DS BULERIA. Repositorio Institucional de la Universidad de León
RD 26-may-2024