Single cell RNA sequencing (scRNA-seq) of human embryoid body (day 4 of culture)

Human Primordial Germ Cell (PGC)-like cells (PGCLC) were specified in vitro from naive-like hiPS cell lines. To evaluate the first steps of PGCLC differentiation, we performed scRNA-seq (droplet-based technology, 10x) on cell suspension from embryoid body (EB) obtained after EB aggregation followed by 4 days of culture (EBd4) with cocktail of cytokines (BMP4, LIF, SCF, EGF and ROCK inhibitor) supplemented with 0.1 mM 2-mercaptoethanol and 15% KSR. In this study, two pools of embryoid bodies (EBD4_A, EBD4_B) were cultured independently for for further encapsulation and sequencing.