Department of Animal Sciences, Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University, Rehovot, Israel.
PLoS One. 2022 Oct 26;17(10):e0276642. doi: 10.1371/journal.pone.0276642. eCollection 2022.
The time-lapse system is a non-invasive method that enables a continuous evaluation through embryo development. Here, we examined the association between the morphokinetics of the developing embryo and the transcriptomic profile of the formed blastocysts. Bovine oocytes were matured and fertilized in vitro; then, the putative zygotes were cultured in an incubator equipped with a time-lapse system. Based on the first-cleavage pattern, embryos were categorized as normal or abnormal (68.5±2.2 and 31.6±2.3%, respectively; P<0.001). A cleaved embryo was defined as normal when it first cleaved into two equal blastomeres; it was classified as synchronous or asynchronous according to its subsequent cleavages. An abnormal pattern was defined as direct, unequal, or reverse cleavage. Direct cleavage was classified as division from one cell directly into three or more blastomeres; unequal cleavage was classified as division that resulted in asymmetrically sized blastomeres; and reverse cleavage of the first division was classified as reduced number of blastomeres from two to one. Of the normally cleaving embryos, 60.2±3.1% underwent synchronous cleavage into 4, 8, and 16 blastomeres, and 39.7±3.1% cleaved asynchronously (P<0.001). The blastocyte formation rate was lower for the synchronously vs. the asynchronously cleaved embryos (P<0.03). The abnormally cleaved embryos showed low competence to develop to blastocysts, relative to the normally cleaved embryos (P<0.001). Microarray analysis revealed 895 and 643 differentially expressed genes in blastocysts that developed from synchronously and asynchronously cleaved embryos, respectively, relative to those that developed from directly cleaved embryos. The genes were related to the cell cycle, cell differentiation, metabolism, and apoptosis. About 180 differentially expressed genes were found between the synchronously vs. the asynchronously cleaved embryos, related to metabolism and the apoptosis mechanism. We provide the first evidence indicating that an embryo's morphokinetics is associated with the transcriptome profile of the derived blastocyst, which might be practically relevant for the embryo transfer program.
时移系统是一种非侵入性方法,可以通过胚胎发育进行连续评估。在这里,我们研究了胚胎发育的形态动力学与形成的囊胚转录组特征之间的关联。牛卵母细胞在体外成熟和受精;然后,将假定的受精卵在配备有时间推移系统的孵育箱中培养。根据第一次卵裂模式,胚胎分为正常或异常(分别为 68.5±2.2%和 31.6±2.3%;P<0.001)。当一个分裂的胚胎第一次分裂成两个相等的卵裂球时,它被定义为正常;根据随后的分裂,它被分类为同步或异步。异常模式定义为直接、不等或反向分裂。直接分裂被定义为一个细胞直接分裂成三个或更多的卵裂球;不等分裂被定义为分裂导致卵裂球大小不对称;第一次分裂的反向分裂被定义为从两个减少到一个卵裂球的数量。在正常分裂的胚胎中,60.2±3.1%同步分裂成 4、8 和 16 个卵裂球,39.7±3.1%异步分裂(P<0.001)。同步分裂的胚胎囊胚形成率低于异步分裂的胚胎(P<0.03)。与正常分裂的胚胎相比,异常分裂的胚胎发育成囊胚的能力较低(P<0.001)。微阵列分析显示,与直接分裂的胚胎相比,同步和异步分裂的胚胎发育成囊胚的胚胎中分别有 895 个和 643 个差异表达基因。这些基因与细胞周期、细胞分化、代谢和凋亡有关。在同步分裂的胚胎与异步分裂的胚胎之间发现了约 180 个差异表达基因,与代谢和凋亡机制有关。我们提供的第一个证据表明,胚胎的形态动力学与衍生囊胚的转录组特征相关,这可能对胚胎移植计划具有实际意义。