Phang C H
Neuropeptide Laboratory, Institute of Molecular and Cell Biology, National University of Singapore, Republic of Singapore.
Methods Mol Biol. 1993;18:265-8. doi: 10.1385/0-89603-245-0:265.
Microinjected eggs are normally incubated overnight in Ml6 microdrop culture at 37°C in CO₂ until 1400 h. By this time the eggs will have developed as far as the two-cell stage. The eggs are then implanted into surrogate mothers. As the membrane covering the oviduct of the rat is much tougher than the mouse, we normally cut it with a very fine pair of scissors rather than tearing it. For a female rat that had been successfully induced into pseudopregnancy, the infundibulum will be swollen and the opening of the infundibulum can be clearly seen under the 10-20X magnification of the dissecting microscope.
显微注射的卵通常在含二氧化碳的37°C的Ml6微滴培养液中过夜培养,直至1400时。此时,卵将发育到二细胞期。然后将卵植入代孕母鼠体内。由于覆盖大鼠输卵管的膜比小鼠的坚韧得多,我们通常用一把非常精细的剪刀剪开它,而不是撕开它。对于已成功诱导假孕的雌性大鼠,漏斗部会肿胀,在解剖显微镜10 - 20倍的放大倍数下可以清楚地看到漏斗部的开口。