I. Crespo Félez, S. Pardal Higuera, M. Mata Campuzano, A. Castañeda, Jeffrey J. Morrell, Felipe Martínez-Pastor
Selection of the optimal spermatozoa population is a necessary step in reproductive techniques, and it increases the quality and resilience of spermatozoa. Androcoll is a single-layer selection colloid that has been adapted to many species. We tested Androcoll-O-S (for sheep and small volumes) for selecting cryopreserved spermatozoa from red deer. Samples were collected from the cauda epididymis, frozen (20% egg yolk, 8% glycerol) and selected post-thawing. We tested two sample volumes, 150 and 300 μL, and two RCF, 300 and 600×g (20 min), in a 1-mL Androcoll volume. Analysis (flow cytometry, YO-PRO-1/propidium iodide/Mitotracker deep red) of selected cells (pellet) resulted in improved viability (87.3%±10.9), mitochondrial activity (86.5%±5.7) and apoptotic ratio (4.8%±2.2) than the unselected control (61.4%±10.8, 56.1%±8.2, 8.9%±2.7), with 300×g achieving slightly higher results for mitochondrial activity. Low quality spermatozoa (48.4%±9, 39.4%±10.2, 9.9%±5.1) were retained in the interphase. Recovery (selected/unselected cells, overall 29.1%±10.9) showed an interaction between volume and RCF, with increasing RCF not improving recovery when using 300 μL at 600×g (26.2%±10.1 vs. 34.3%±13.5 for 150 μL), maybe because a higher number of cells at these RCF could impair that high-quality cells enter the colloid. Androcoll-O-S is convenient and effective for selecting high-quality spermatozoa from thawed red deer.
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