Development to term of sheep embryos reconstructed after inner cell mass/trophoblast exchange

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  • LOI Pasqualino
    Laboratory of Embryology, Faculty of Veterinary Medicine, University of Teramo, 64100 Teramo, Italy
  • GALLI Cesare
    Avantea srl., Laboratorio di Tecnologie della Riproduzione, Cremona, Italy
  • LAZZARI Giovanna
    Avantea srl., Laboratorio di Tecnologie della Riproduzione, Cremona, Italy
  • MATSUKAWA Kazutsugu
    Research and Education Faculty, Multidisciplinary Science Cluster, Life and Environmental Medicine Science Unit, Kochi University, Kochi 783-8502, Japan
  • FULKA, Jr Josef Jr.
    Institute of Animal Science, Prague, Czech Republic
  • GOERITZ Frank
    Leibniz Institute for Zoo & Wildlife Research, Berlin, Germany
  • HILDEBRANDT Thomas B.
    Leibniz Institute for Zoo & Wildlife Research, Berlin, Germany

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Abstract

<p> Here we report in vitro and term development of sheep embryos after the inner cell mass (ICM) from one set of sheep blastocysts were injected into the trophoblast vesicles of another set. We also observed successful in vitro development of chimeric blastocysts made from sheep trophoblast vesicles injected with bovine ICM. First, we dissected ICMs from 35 sheep blastocysts using a stainless steel microblade and injected them into 29 re-expanded sheep trophoblastic vesicles. Of the 25 successfully micromanipulated trophoblastic vesicles, 15 (51.7%) re-expanded normally and showed proper ICM integration. The seven most well reconstructed embryos were transferred for development to term. Three ewes receiving manipulated blastocysts were pregnant at day 45 (42.8%), and all delivered normal offspring (singletons, two females and one male, average weight: 3.54 ± 0.358 kg). Next, we monitored in vitro development of sheep trophoblasts injected with bovine ICMs. Of 17 injected trophoblastic vesicles, 10 (58.8%) re-expanded after 4 h in culture, and four (40%) exhibited integrated bovine ICM. Our results indicate that ICM/trophoblast exchange is feasible, allowing full term development with satisfactory lambing rate. Therefore, ICM exchange is a promising approach for endangered species conservation.</p>

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