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Oocyte activation procedures and influence of serum on porcine oocyte maturation and subsequent parthenogenetic and nuclear transfer embryo development

Published online by Cambridge University Press:  01 November 2008

E. García-Mengual ,
J. Alfonso ,
I. Salvador ,
C-C. Duque  and
M-A. Silvestre
E. García-Mengual
Affiliation:
Centro de Tecnología Animal. Instituto Valenciano de Investigaciones Agrarias (CITA-IVIA), Segorbe, Castellón, Spain
J. Alfonso
Affiliation:
Instituto de Medicina Reproductiva (IMER), Valencia, Spain.
I. Salvador
Affiliation:
Centro de Tecnología Animal. Instituto Valenciano de Investigaciones Agrarias (CITA-IVIA), Segorbe, Castellón, Spain
C-C. Duque
Affiliation:
Unidad de Reproducción Asistida Humana. H.U. La Fe de Valencia, Valencia, Spain.
M-A. Silvestre*
Affiliation:
Centro de Tecnología Animal. Instituto Valenciano de Investigaciones Agrarias (CITA-IVIA). Apdo 187, Pol. La Esperanza n°100, 12400, Segorbe, Castellón, Spain Centro de Tecnología Animal. Instituto Valenciano de Investigaciones Agrarias (CITA-IVIA), Segorbe, Castellón, Spain
*
All correspondence to: M-A Silvestre, Centro de Tecnología Animal. Instituto Valenciano de Investigaciones Agrarias (CITA-IVIA). Apdo 187, Pol. La Esperanza n°100, 12400, Segorbe, Castellón, Spain; Tel: +34964712115, Fax: +34964710218, e-mail: masilver@ivia.es.
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Summary

The viability of SCNT embryos is poor, with an extremely low cloned piglet production rate. In the present work, we studied the effect of three activation protocols based on ionomycin treatment (5 μM ionomycin for 5 min and incubated in 2 mM 6-DMAP for 3.5 h) or electric stimuli (two square wave electrical DC pulses of 1.2 kV/cm for 30 μs) combined or not with 6-DMAP on parthenogenetic embryo development. Oocytes activated by ionomycin plus 6-DMAP showed lower cleavage (47.2 vs. 78.5–81.5; p < 0.05) and blastocyst rates (11.3 vs. 29.2–32.1; p < 0.05) than those activated by electrical and electrical plus 6-DMAP treatments. Also, we studied the effect of addition of serum to maturation medium (0% vs. 10%) on nuclear maturation and further parthenogenetic and SCNT embryo development. We observed in the parthenogenetic embryos that cleavage rates in the serum-free group were significantly higher than in the serum-supplemented group (81.8 vs. 69.6% respectively; p < 0.05), although these differences were not detected in blastocyst rates or blastocyst nuclei numbers. Regarding SCNT embryos, no significant differences were observed in cleavage or blastocyst rates between different experimental groups of SCNT embryos. In conclusion, electrical pulse followed or not by 6-DMAP was found to be an efficient procedure to artificially activate MII porcine oocytes. Moreover, the addition of serum to oocyte maturation media did not seem to improve parthenogenetic or SCNT porcine embryo development.

Keywords

Nuclear transferOocyte maturationParthenogenetic activationPorcineSerum
Type
Research Article
Information
Zygote , Volume 16 , Issue 4 , November 2008 , pp. 279 - 284
Copyright
Copyright © Cambridge University Press 2008

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