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Comparison of male chimeric mice generated from microinjection of JM8.N4 embryonic stem cells into C57BL/6J and C57BL/6NTac blastocysts |
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| Authors: | Thomas J Fielder Charles S Yi Juliet Masumi Katrina G Waymire Hsiao-Wen Chen Shuling Wang Kai-Xuan Shi Douglas C Wallace Grant R MacGregor |
| Institution: | 1. UCI Transgenic Mouse Facility, University Laboratory Animal Resources, Office of Research, University of California, Irvine, Irvine, CA, 92697-1140, USA 2. Department of Developmental and Cell Biology, School of Biological Sciences, University of California, Irvine, Irvine, CA, 92697-2300, USA 3. Center for Molecular and Mitochondrial Medicine and Genetics, University of California, Irvine, Irvine, CA, 92697-3490, USA 4. Department of Biological Chemistry, School of Medicine, University of California, Irvine, Irvine, CA, 92697-1700, USA 5. Department of Ecology and Evolutionary Biology, School of Biological Sciences, University of California, Irvine, Irvine, CA, 92697-2525, USA 6. Center for Mitochondrial and Epigenomic Medicine, Children??s Hospital of Philadelphia Research Institute, Philadelphia, PA, 19104, USA 7. Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, 92697-2300, USA |
| Abstract: | To identify ways to improve the efficiency of generating chimeric mice via microinjection of blastocysts with ES cells, we compared production and performance of ES-cell derived chimeric mice using blastocysts from two closely related and commonly used sub-strains of C57BL/6. Chimeras were produced by injection of the same JM8.N4 (C57BL/6NTac) derived ES cell line into blastocysts of mixed sex from either C57BL/6J (B6J) or C57BL/6NTac (B6NTac) mice. Similar efficiency of production and sex-conversion of chimeric animals was observed with each strain of blastocyst. However, B6J chimeric males had fewer developmental abnormalities involving urogenital and reproductive tissues (1/12, 8?%) compared with B6NTac chimeric males (7/9, 78?%). The low sample size did not permit determination of statistical significance for many parameters. However, in each category analyzed the B6J-derived chimeric males performed as well, or better, than their B6NTac counterparts. Twelve of 14 (86?%) B6J male chimeras were fertile compared with 6 of 11 (55?%) B6NTac male chimeras. Ten of 12 (83?%) B6J chimeric males sired more than 1 litter compared with only 3 of 6 (50?%) B6NTac chimeras. B6J male chimeras produced more litters per productive mating (3.42?±?1.73, n?=?12) compared to B6NTac chimeras (2.17?±?1.33, n?=?6). Finally, a greater ratio of germline transmitting chimeric males was obtained using B6J blastocysts (9/14; 64?%) compared with chimeras produced using B6NTac blastocysts (4/11; 36?%). Use of B6J host blastocysts for microinjection of ES cells may offer improvements over blastocysts from B6NTac and possibly other sub-strains of C57BL/6 mice. |
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