Production of cloned bovine embryos by somatic cell transfer into enucleated zona-free oocytes |
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Authors: | G P Malenko M I Prokof’ev M V Pinyugina T A Antipova M N Mezina Yu M Bukreev |
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Institution: | (1) Biotechcenter Department, Afanasiev Research Institute of Fur Bearing Animals and Rabbits, Russian Academy of Agricultural Sciences, Administrativnaya ul. 2, Gorki Leninskie, Moscow oblast, 142712, Russia;(2) JSC Scientific and Technical Center Novenergo, Parkovaya ul. 12, Krasnoznamensk, Moscow oblast, 143090, Russia |
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Abstract: | Cloned bovine embryos were produced at the blastocyst stage. Prior to enucleation, oocytes were freed from the zona pellucida. Fibroblasts isolated from the bovine fetus were used as nuclear donors. Pairs of fetal fibroblasts and enucleated oocytes (cytoplasts) were glued in phytohemagglutinin solution under a binocular microscope. The subsequent electrofusion of 39 fetal fibroblast-cytoplast pairs yielded 36 reconstructed one-cell embryos (92.3%). After culturing in synthetic oviduct fluid for 7.5 days, seven cloned embryos developed to the blastocyst stage (19.4%) and six blastocysts were considered fit for transplantation. The applied technique of bovine embryo growth allowed 31.1% zona-free oocytes parthenogenetically activated by to reach the blastocyst stage. |
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