New Methods for Selecting Stallion Spermatozoa for Assisted Reproduction |
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| Authors: | Silvia Colleoni PhD Irina Lagutina PhD Giovanna Lazzari DVM PhD Heriberto Rodriguez-Martinez Prof DVM Cesare Galli Prof DVM Jane M Morrell BVetMed PhD |
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| Institution: | aAVANTEA, Laboratorio di Tecnologie della Riproduzione, Cremona, Italy;bDepartment of Clinical and Experimental Medicine, Linköping University, Linköping, Sweden;cDipartimento Clinico Veterinario, Università di Bologna, Bologna, Italy;dDivision of Reproduction, Swedish University of Agricultural Sciences, Uppsala, Sweden |
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| Abstract: | Improved sperm selection techniques are needed to increase the efficiency of equine-assisted reproduction. Single layer centrifugation (SLC) of spermatozoa has been shown to improve the quality of stallion sperm samples. In this study, the functionality of selected stallion spermatozoa was tested by intracytoplasmic sperm injection of equine oocytes after selection by SLC through Androcoll-E or by discontinuous density gradient centrifugation (DGC) through Redigrad and Tyrode's medium with added albumin, lactate, and pyruvate. The mean cleavage rates of the injected oocytes from SLC- and DGC-selected spermatozoa were 67% and 66%, respectively, whereas the proportion of blastocysts developing from cleaved oocytes was 28% and 22%, respectively (P > .05, not significant). An incidental finding was that there was a tendency for SLC-selected spermatozoa to have a higher percentage of spermatozoa with normal morphology than DGC (70% ± 22% vs. 58% ± 38%) and for more blastocysts to be obtained from subfertile ejaculates (21 19.6%] vs. 15 14.4%], respectively). In further experiments, stallion spermatozoa bound to hyaluronan, although binding may depend on the semen extender and sperm treatment as well as incubation time. In conclusion, SLC-selected stallion spermatozoa function normally when injected into oocytes. SLC may potentially be better than DGC at selecting spermatozoa from subfertile ejaculates, but this effect needs rigorous investigation with a much larger sample size. Use of the hyaluronan-binding assay for assessing the potential fertility of stallion spermatozoa may be useful but requires further evaluation. |
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| Keywords: | ICSI Stallion spermatozoa Hyaluronan binding Androcoll-E Single layer centrifugation |
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