Faculty of Veterinary Medicine, Ghent University, B-9820 Merelbeke, Belgium.
Anal Biochem. 2011 Sep 15;416(2):228-30. doi: 10.1016/j.ab.2011.05.033. Epub 2011 May 27.
Several approaches have been described for differential staining of blastocysts, but these methods are often time-consuming and unreliable. Here we describe a method for simultaneous differential staining and detection of apoptosis. The differential staining is based on the transcription factor CDX2 which is localized in the nucleus of trophectoderm (TE) cells but absent in the inner cell mass (ICM). Apoptosis is detected by staining of active caspase-3, a key player in several apoptotic pathways. This new approach represents a robust method for quantifying simultaneously ICM/TE ratio and apoptotic cell ratio in bovine, murine, porcine, and human blastocysts.
已经描述了几种用于囊胚差异染色的方法，但这些方法通常耗时且不可靠。在这里，我们描述了一种用于同时进行差异染色和检测凋亡的方法。这种差异染色基于转录因子 CDX2，它定位于滋养外胚层 (TE) 细胞的核内，但不存在于内细胞团 (ICM) 中。凋亡通过染色活性半胱氨酸蛋白酶-3 来检测，该酶是几种凋亡途径中的关键因子。这种新方法是一种强大的方法，可用于定量牛、鼠、猪和人囊胚中的 ICM/TE 比和凋亡细胞比。
