Impact of the MACS on elimination of apoptotic spermatozoa from rabbit ejaculates

Abstract

The objective of this study was to assess the effectiveness of the MACS technique used for the elimination of apoptotic rabbit spermatozoa from heterospermic pool (Experiment 1) as well as from the ejaculates of individual bucks (Experiment 2). The semen samples from control (untreated) and magnetically separated spermatozoa (in both E1 and E2) were evaluated by fluorescence analysis using Annexin-V-FLUOS Staining Kit (Roche Slovakia, Slovak Republic). Superparamagnetic microbeads conjugated with annexin V eliminated spermatozoa with externalized phosphatidylserine via MACS. MACS separation of spermatozoa yields two fractions: the annexin V-negative (AnV- ) and the annexin V-positive (AnV+ ). The number of AnV+ sperm was significantly lower (P<0.001) in the AnV- fractions than in the AnV+ fractions (in both E1 and E2). Our observations indicate that MACS technique could be an adequate method for the elimination of apoptotic spermatozoa with externalized phosphatidylserine from the rabbit ejaculates. However, further experiments are required in order to prove this suggestion.