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KMID : 1023620040280040257
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Reproductive and Developmental Biology
2004 Volume.28 No. 4 p.257 ~ p.260
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Polscope-Assisted Enucleation for Nuclear Transfer in Mice
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Won Ji-Young
Kang Jee-Hyun
Shim Ho-Sup
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Abstract
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Efficiency of somatic cell nuclear transfer was investigated in mice. First, oocyte activation was induced by SrCl₂, and the rate of development was compared with embryos from normal fertilization. Although more than one half of SrCl₂-treated oocytes developed to blastocysts (146/262, 55.7%), the rate of blastocyst formation was significantly lower than normal fertilization controls (59/79, 74.6%). Second, enucleation of oocytes was performed using Polscope that enables non-invasive visualization of metaphase spindles. Such approach could not only avoid damage of oocytes during an exposure to UV light often employed in conventional enucleation procedures, but could also assure the removal of nuclei from all oocytes operated because of monitoring the location of spindles during an entire process of enucleation. Morphologically normal blastocysts were obtained from the transfer of cumulus cell nuclei into enucleated oocytes. However, the rate of development into the blastocyst stage was still low (4/93, 4.3%). This reflects that the nuclear transfer procedure used in this study was not sufficiently optimized, and other factors may also impact greatly the efficiency of nuclear transfer. Including an induction of oocyte activation and method of enucleation tested in this study, a lot more elements are remained to be optimized to improve the efficiency of somatic cell nuclear transfer in mice.
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KEYWORD
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Polscope, Enucleation, Nuclear transfer, Mouse
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