Valencia Node of The National Stem Cell Bank, Centro de Investigación Príncipe Felipe, Valencia, Spain.
PLoS One. 2010 Oct 26;5(10):e13615. doi: 10.1371/journal.pone.0013615.
Blastomere fate and embryonic genome activation (EGA) during human embryonic development are unsolved areas of high scientific and clinical interest. Forty-nine blastomeres from 5- to 8-cell human embryos have been investigated following an efficient single-cell cDNA amplification protocol to provide a template for high-density microarray analysis. The previously described markers, characteristic of Inner Cell Mass (ICM) (n = 120), stemness (n = 190) and Trophectoderm (TE) (n = 45), were analyzed, and a housekeeping pattern of 46 genes was established. All the human blastomeres from the 5- to 8-cell stage embryo displayed a common gene expression pattern corresponding to ICM markers (e.g., DDX3, FOXD3, LEFTY1, MYC, NANOG, POU5F1), stemness (e.g., POU5F1, DNMT3B, GABRB3, SOX2, ZFP42, TERT), and TE markers (e.g., GATA6, EOMES, CDX2, LHCGR). The EGA profile was also investigated between the 5-6- and 8-cell stage embryos, and compared to the blastocyst stage. Known genes (n = 92) such as depleted maternal transcripts (e.g., CCNA1, CCNB1, DPPA2) and embryo-specific activation (e.g., POU5F1, CDH1, DPPA4), as well as novel genes, were confirmed. In summary, the global single-cell cDNA amplification microarray analysis of the 5- to 8-cell stage human embryos reveals that blastomere fate is not committed to ICM or TE. Finally, new EGA features in human embryogenesis are presented.
人类胚胎发育过程中的卵裂球命运和胚胎基因组激活（EGA）是具有高度科学和临床兴趣的未解决领域。通过高效的单细胞 cDNA 扩增方案，对 5-8 细胞期人类胚胎的 49 个卵裂球进行了研究，为高密度微阵列分析提供了模板。分析了先前描述的内细胞团（ICM）标志物（n = 120）、干性标志物（n = 190）和滋养外胚层（TE）标志物（n = 45），并建立了 46 个管家基因的表达模式。5-8 细胞期胚胎的所有人类卵裂球均表现出与 ICM 标志物（例如 DDX3、FOXD3、LEFTY1、MYC、NANOG、POU5F1）、干性标志物（例如 POU5F1、DNMT3B、GABRB3、SOX2、ZFP42、TERT）和 TE 标志物（例如 GATA6、EOMES、CDX2、LHCGR）相对应的共同基因表达模式。还研究了 5-6 细胞期和 8 细胞期胚胎之间的 EGA 特征，并与囊胚期进行了比较。确认了已知基因（n = 92），例如耗尽的母体转录物（例如 CCNA1、CCNB1、DPPA2）和胚胎特异性激活基因（例如 POU5F1、CDH1、DPPA4），以及新的基因。总之，5-8 细胞期人类胚胎的全局单细胞 cDNA 扩增微阵列分析表明，卵裂球命运尚未确定为 ICM 或 TE。最后，提出了人类胚胎发生中的新的 EGA 特征。
