Podocalyxin-Like Protein 1 Regulates Extended and Primed Pluripotency through the Cholesterol Biosynthesis Pathway

We utilized scRNA-seq to study the human-mouse chimera transcriptomic regulation at peri-implantation embryo stage. Single-cell RNA sequencing revealed PODXL overexpression boosted chimerism between human cells in mouse host embryos and the potential cell-cell communication interactions in human-mouse chimeras. Potential cell-cell communications were identified to optimize cell-cell contact interaction for reducing the xenogeneic barrier in the interspecies chimeras in future studies. We generated Tet-on-PODXL hEPSC lines that stably expressed tdTomato (tdT) florescence protein. We generated human-mouse chimeric embryos by incorporating one 8C embryo with ~8-10 cells of Dox-treated or untreated control hEPSCs using an aggregation method. The embryos were cultured for additional ~37 hours to the late blastocyst stage.