Abstract

The aim of our work was to analyse the expression pattern of embryonic stem cell specificmarkers in rabbit embryos and epiblast cells. There are well-known genes that direct pluripotency inthe mouse, such as Oct4, Nanog, but their expression pattern during the rabbit embryonicdevelopment is not known yet. Using culture medium supplemented with two inhibitors [(2i:PD032590-(PD), CHIR99021-(CH); embryonic stem cell (ESC) lines could be derived efficientlyfrom mouse and rat embryos. We cultured the rabbit embryos from eight-cell-stage in RDH, RDH+2iculture medium until the blastocyst stage. The expression level of epiblast specific Oct4 and Nanog,trophoblast specific Cdx2 and hypoblast specific Gata4 and Gata6 were examined in blastocysts andattached embryoblasts. The expression level of Oct4 and Nanog was significantly higher inblastocysts, cultured in RDH+2i culture medium, but there was no significant difference in CDX2,GATA4, GATA6 expression on mRNA and protein level, compared to embryos cultured in RDHmedium. We hope that we can maintain the pluripotency of the epiblast derived stem cells in 2icontaining medium during long term cultivation.

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