Single cell transciptional profiling of a hESC 3D extra-embryonic amnoid model (PGA - post-gastrulation amnoid)

The goal of this study was to carry out a comprehensive molecular characterization of transcriptional profile and cell types formed during amniogenesis at the single-cell level. We further explored the transcriptional profiles and cell types formed over time when the extra-embryonic cells were mixed with (primed) hESC representing cells of the inner cell mass epiblast. Overall design: scRNA-seq was performed on cultured hESC line H1 grown in mTESR + matrigel. For induction of extraembryonic lineages, cells were treated with BMP4 (24h) and CHIR99021 (24h). For formation of co-culture samples, WT hESC grown in mTESR were mixed with extraembryonic-induced cells. Samples were taken at 0, 1, 3, 5 and 8 days for scRNA-seq analysis. Two biological replicates per condition were multiplexed using Cell Multiplexing Oligos (CMO) (10x Genomics 3' Cellplex kit set A (PN-1000261)) and sequenced together.