Functional genomics of 5- to 8-cell stage human embryos by blastomere single-cell cDNA analysis.
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.
Valencia Node of The National Stem Cell Bank, Centro de Investigación Príncipe Felipe, Valencia, Spain.
Amparo
A
Galán
David
D
Montaner
M Eugenia
ME
Póo
Diana
D
Valbuena
Verónica
V
Ruiz
Cristóbal
C
Aguilar
Joaquín
J
Dopazo
Carlos
C
Simón
D001757:Blastomeres / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics*; D020869:Gene Expression Profiling; D023281:Genomics; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis
D018076:DNA, Complementary