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KMID : 0900920010250010071
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Korean journal of Animal Reproduction
2001 Volume.25 No. 1 p.71 ~ p.77
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Factors Affecting the Productivity of Germ-line Chimeras from Jl Embryonic Stem Cells
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Kim S.-U.
Koo B.-S.
Jeong S.
Lee T.-H.
Yu S.-L.
Nam Y.-I.
Kim J.-L.
Hyun B.-H.
Shin H.-S.
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Abstract
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This experiment was designed to improve the production efficiency of germ-line chimeric mice from phospholipase C (PLC)-beta3 or peroxiredoxin (Prx) E -targeted (Delta) ES cells by the investigating the manipulation conditions and characteristics of Jl ES cells. Four targeting clones were isolated to investigate the karyotypical and morphological stability prior to injection. All clones (DeltaPLCbeta-3 C3, DeltaPrx II C3, C10 and I5) showed more than 80% euploidism, however, most of DeltaPLCbeta-3 C3 clones were extensively differentiated compared to the other clones. Nine of 13 DeltaPrx II chimeras appeared to have at least 80% chimerism, whereas DeltaPLCbeta-3 C3 chimeras had 20% chimerism at most. Therefore, the morphological stability of ES cells under stable euploidism might mainly affect the production rate of high-coat chimeric mice. To increase the collection rate of injectable blastocysts (IBs), 5 to 10 week -aged C57BL/6J female mice were sacrificed at 3.5 days post-coitum. Ten week-aged mice were the most optimal IB donors by showing the highest collection rate (2.94/mouse) of injectable blastocysts without increase of non-injectable embryos (0.29/mouse). Foster mothers might be another factor because ICR x C57BL/6J F1 foster mother showed more increased productivity in litter size (2.8 vs. 5.6) and chimera (0 vs. 35.3%) than those of ICR foster mothers. In conclusion, the efficient production of germ-line chimeras mainly depends on the maintenance of ES cell morphology during targeting procedure, and the establishment of manipulation conditions might be a key point to maximize it.
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KEYWORD
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Embryonic stem cells, Chimerism, Germ-line chimera, Morphology, Karyotype
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