Malenko G P, Prokof'ev M I, Piniugina M V, Antipova T A, Mezina M N, Bukreev Iu M
Izv Akad Nauk Ser Biol. 2006 May-Jun(3):284-91.
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.
克隆牛胚胎在囊胚阶段产生。去核前,将卵母细胞从透明带中取出。从牛胎儿分离出的成纤维细胞用作核供体。在双目显微镜下,将成对的胎儿成纤维细胞和去核卵母细胞(细胞质体)在植物血凝素溶液中黏合。39对胎儿成纤维细胞 - 细胞质体对随后的电融合产生了36个重构的单细胞胚胎(92.3%)。在合成输卵管液中培养7.5天后,7个克隆胚胎发育到囊胚阶段(19.4%),6个囊胚被认为适合移植。所应用的牛胚胎生长技术使31.1%经孤雌激活的无透明带卵母细胞能够发育到囊胚阶段。