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KMID : 1161520140180050324
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Animal Cells and Systems
2014 Volume.18 No. 5 p.324 ~ p.332
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Production of cloned mice by aggregation of tetraploid embryo
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Sim Bo-Woong
Min Kwan-Sik
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Abstract
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Mouse chimeras can also successfully be produced using tetraploid host embryos. This study was conducted to optimize the efficiency of cloning and to produce cloned mice using tetraploid host embryos. Six hours of activation with strontium (SrCl2) was optimal for somatic cell nuclear transfer (SCNT) embryos. Cytochalasin B (CB) concentration (5 μg/ml) during enucleation was evaluated in the efficiency of implantation sites and fetus offspring. Continuous exposure to 5?50 nM trichostatin A (TSA), a histone-deacetylase inhibitor (HDACi), for 10 h is recommended for production of clone mice. Aggregated SCNTs were constructed by aggregation of SCNT embryos with tetraploid embryos to reduce epigenetic errors in the placenta. The pregnancy and implantation rates of aggregated SCNT were significantly higher than those of SCNT alone. The full-term developmental rate of aggregated embryos was also higher than that of SCNT (3.57 vs. 1.16). The placental weight of SCNT clones was significantly higher than that from in vitro fertilization (IVF). However, the placenta weight of aggregated SCNT clones was nearly the same as that of embryos in the IVF group. The placentas of SCNT-only clones appeared to have the hyperplastic histology typical of mouse clones. We produced a total of 36 clone mice, including nine heads derived from aggregated SCNT. One-half of clones derived from aggregated SCNT survived to adulthood, and 14-clones derived from SCNT grew into healthy adults. The aggregated SCNT method was useful for significantly reducing the placental weight of cloned mice and improving the efficiency of SCNT.
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KEYWORD
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cloning, mice, aggregation, tetraploid, placenta
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