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Published-Ahead-of-Print November 22, 2005, DOI:10.2164/jandrol.05110

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Influence of sperm pretreatment on the efficiency of Intracytoplasmic Sperm Injection (ICSI) in pigs

Empar Garcia-Rosello , Carmen Matas , Sebastian Canovas , Pedro N. Moreira , Joaquin Gadea , and Pilar Coy

The purpose of this study was to determine the influence of sperm pretreatment on the efficiency of Intracytoplasmic Sperm Injection (ICSI) in pigs. This was done by examining the effect of 1) the conservation method (fresh vs frozen); 2) the sperm treatment pre-injection (resuspension in DPBS vs selection by a Percoll® gradient) and 3) the acrosomal and live or dead status of the spermatozoa (by incubation with or without calcium ionophore 1µM and 5µM). In vitro matured porcine oocytes were injected with treated spermatozoa according to each experiment. All the experiments were done with non-artificially activated oocytes. The percentages of activation and cleavage were higher (68% vs. 43% and 63% vs.43%, respectively, p<0.05) in oocytes injected with fresh versus frozen spermatozoa. The DPBS treatment allowed higher cleavage proportions than the Percoll® treatment (p<0.05). Moreover, a boar effect was observed in the percentage of developing blastocysts. None of the studied parameters was affected by the acrosomal or the live or dead status of the spermatozoa injected. In conclusion, the use of fresh semen is recommended for porcine ICSI, as well as careful selection of the boar; Percoll® treatment is only recommended for poor quality samples or removing toxic agents, and no exogenous form of activation or induction of the acrosome reaction is necessary for porcine oocytes to develop a male pronucleus and cleave up to the 2 cell stage after ICSI, although experimental conditions to reach the blastocyst stage need to be investigated further.




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M. A. Silvestre, J. Alfonso, E. Garcia-Mengual, I. Salvador, C. C. Duque, and I. Molina
Effect of recombinant human follicle-stimulating hormone and luteinizing hormone on in vitro maturation of porcine oocytes evaluated by the subsequent in vitro development of embryos obtained by in vitro fertilization, intracytoplasmic sperm injection, or parthenogenetic activation
J Anim Sci, May 1, 2007; 85(5): 1156 - 1160.
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S. Canovas, P. Coy, and E. Gomez
First Steps in the Development of a Functional Assay for Human Sperm Using Pig Oocytes
J Androl, March 1, 2007; 28(2): 273 - 281.
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