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Influence of oocyte selection, activation with a zinc chelator and inhibition of histone deacetylases on cloned porcine embryo and chemically activated oocytes development

Published online by Cambridge University Press:  14 April 2020

Felipe L. Ongaratto*
Affiliation:
Recombinetics Inc., Saint Paul, Minnesota, USA
Paula Rodriguez-Villamil
Affiliation:
Recombinetics Inc., Saint Paul, Minnesota, USA
Marcelo Bertolini
Affiliation:
Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
Daniel F. Carlson
Affiliation:
Recombinetics Inc., Saint Paul, Minnesota, USA
*
Author for correspondence: Felipe L. Ongaratto. Recombinetics Inc., 1246 University Ave W, Saint Paul, MN55104, USA. Tel: +1 612 727 2000. E-mail: felipe.ongaratto@recombinetics.com

Summary

The aim of this study was to evaluate the effects of alternative protocols to improve oocyte selection, embryo activation and genomic reprogramming on in vitro development of porcine embryos cloned by somatic cell nuclear transfer (SCNT). In Experiment 1, in vitro-matured oocytes were selected by exposure to a hyperosmotic sucrose solution prior to micromanipulation. In Experiment 2, an alternative chemical activation protocol using a zinc chelator as an adjuvant (ionomycin + N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) + N-6-dimethylaminopurine (6-DMAP)) was compared with a standard protocol (ionomycin + 6-DMAP) for the activation of porcine oocytes or SCNT embryos. In Experiment 3, presumptive cloned zygotes were incubated after chemical activation in a histone deacetylase inhibitor (Scriptaid) for 15 h, with the evaluation of embryo yield and total cell number in day 7 blastocysts. In Experiment 1, cleavage rates tended to be higher in sucrose-treated oocytes than controls (123/199, 61.8% vs. 119/222, 53.6%, respectively); however, blastocyst rates were similar between groups. In Experiment 2, cleavage rates were higher in zygotes treated with TPEN than controls but no difference in blastocyst rates between groups occurred. For Experiment 3, the exposure to Scriptaid did not improve embryo development after cloning. Nevertheless, the total number of cells was higher in cloned zygotes treated with Scriptaid than SCNT controls. In conclusion, oocyte selection by sucrose as well as treatments with zinc chelator and an inhibitor of histone deacetylases did not significantly improve blastocyst yield in cloned and parthenotes. However, the histone deacetylases inhibitor produced a significant improvement in the blastocyst quality.

Type
Research Article
Copyright
© Cambridge University Press 2020

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