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2 May 2012 Equine Cloning: In Vitro and In Vivo Development of Aggregated Embryos
Andrés Gambini, Javier Jarazo, Ramiro Olivera, Daniel F. Salamone
Author Affiliations +
Abstract

The production of cloned equine embryos remains highly inefficient. Embryo aggregation has not yet been tested in the equine, and it might represent an interesting strategy to improve embryo development. This study evaluated the effect of cloned embryo aggregation on in vitro and in vivo equine embryo development. Zona-free reconstructed embryos were individually cultured in microwells (nonaggregated group) or as 2- or 3-embryo aggregates (aggregated groups). For in vitro development, they were cultured until blastocyst stage and then either fixed for Oct-4 immunocytochemical staining or maintained in in vitro culture where blastocyst expansion was measured daily until Day 17 or the day on which they collapsed. For in vivo assays, Day 7–8 blastocysts were transferred to synchronized mares and resultant vesicles, and cloned embryos were measured by ultrasonography. Embryo aggregation improved blastocyst rates on a per well basis, and aggregation did not imply additional oocytes to obtain blastocysts. Embryo aggregation improved embryo quality, nevertheless it did not affect Day 8 and Day 16 blastocyst Oct-4 expression patterns. Equine cloned blastocysts expanded and increased their cell numbers when they were maintained in in vitro culture, describing a particular pattern of embryo growth that was unexpectedly independent of embryo aggregation, as all embryos reached similar size after Day 7. Early pregnancy rates were higher using blastocysts derived from aggregated embryos, and advanced pregnancies as live healthy foals also resulted from aggregated embryos. These results indicate that the strategy of aggregating embryos can improve their development, supporting the establishment of equine cloned pregnancies.

© 2012 by the Society for the Study of Reproduction, Inc.
Andrés Gambini, Javier Jarazo, Ramiro Olivera, and Daniel F. Salamone "Equine Cloning: In Vitro and In Vivo Development of Aggregated Embryos," Biology of Reproduction 87(1), (2 May 2012). https://doi.org/10.1095/biolreprod.112.098855
Received: 3 January 2012; Accepted: 1 April 2012; Published: 2 May 2012
KEYWORDS
embryo aggregation
equine cloning
Oct-4
pregnancy
zona-free embryo culture
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