Evaluation of novel swim-up approaches on canine sperm progressive motility, concentration, morphology, and acrosome reaction Public

http://ir.library.oregonstate.edu/concern/honors_college_theses/xw42n976w

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  • Introduction: Swim-up has been considered a preferred method for separating non-motile sperm in an ejaculate. However, the long incubation time and equipment necessary for swim-up make this process clinically impractical. It was hypothesized that these problems could be avoided for canine applications with a shorter incubation time at room temperature (RT). The objective was to compare the standard swim-up method (STAN) to this novel, modified method (MOD). In addition, it was hypothesized that incubation with heat-treated canine serum (HTCS) would improve swim-up results compared to incubation with bovine serum albumin (BSA) or fetal calf serum (FCS). Methods: Ejaculates were collected manually from 8 dogs and divided equally into 6 aliquots. Ham’s F-10 supplemented with 10% BSA, FCS or HTCS was overlaid on each semen aliquot. For the STAN, the samples were incubated at 45° angle for 1 hour at 37°C in 5% CO₂. For the MOD, the samples were incubated upright at RT for 5 min. After incubation, 0.5 mL was removed from the top of the sample and progressive motility (PM), sperm concentration (SC), and acrosomal reaction (AR) were evaluated. MOD was compared to STAN via a paired Student's t test and BSA, FCS, and HTCS were compared via a one-way ANOVA Significance was defined as p<0.05. Results: MOD-HTCS and MOD-FCS yielded higher SC compared to the STAN-HTCS and STAN-FCS, respectively (p<0.05), with the same trend observed between MODBSA and STAN-BSA (p=0.05). Irrespective of protein source, MOD always yielded a higher PM and lower AR compared to STAN (p<0.05). MOD-FCS and MOD-BSA had a higher PM compared to MOD-HTCS, and MOD-FCS had a higher PM than MOD-BSA (p<0.05). MOD-FCS had the lowest AR compared to MOD-BSA and MOD-HTCS (p<0.05). Discussion: This modified swim-up method can be easily applied in a clinical setting. Based upon these results, Ham’s F-10 with 10% FCS is superior to either BSA or HTCS when separating the highest concentrations of progressively motile sperm that are not acrosome-reacted.
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