Laminin111-based defined culture promoting self-renewing human pluripotent stem cells exhibiting properties of early post-implantation epiblast cells [LN_hPSC_RRBS]

In our study we sought to develop cultures highly enriched for self-renewing human pluripotent stem cells (hPSCs) with an early post-implantation phenotype, capturing the formative pluripotency phase in vitro. Our results demonstrate that hPSCs cultured on LN111 in defined conditions (LN-hPSCs) generate cultures highly enriched for genetically stable, self-renewing hPSCs exhibiting properties similar to the early-post implantation epiblast, including competence to give rise to the germ cell lineage. To analyze the global transcriptome of LN-hPSCs in comparison with conventional/primed and naive hPSCs, we performed RNA-seq of the three hPSC lines cultured under LN, primed, and naïve conditions. Comparative DNA methylation profiling analysis of reduced representation bisulfite sequencing (RRBS) data for hPSC lines HES-1 (ES01) and HES-2 (ES02) (Reubinoff et al., 2000), cultured under LN, conventional/primed and naïve conditions.