Cell cycle dynamics of human pluripotent stem cells primed for differentiation

Understanding the molecular properties of the cell cycle of human pluripotent stem cells (hPSCs) is critical for effectively promoting differentiation. Here, we use the Fluorescence Ubiquitin Cell Cycle Indicator (FUCCI) system adapted into hPSCs and perform RNA-sequencing on cell cycle sorted hPSCs primed and unprimed for differentiation. Gene expression patterns of signaling factors and developmental regulators change in a cell cycle-specific manner in cells primed for differentiation without altering genes associated with pluripotency. Furthermore, we identify an important role for PI3K signaling in regulating the early transitory states of hPSCs towards differentiation. H9 FUCCI hPSCs were cell-cycle sorted into the early G1, late G1, and SG2M phases. For each phase, cells were treated with 2% DMSO for a 24 hour time course. RNA-seq was performed on the DMSO-treated samples and corresponding controls (2 technical replicates for each cell-cycle phase x DMSO status combination).