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摘要
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 +/- A 1.73, n = 12) compared to B6NTac chimeras (2.17 +/- A 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.
- 出版日期2012-12
