As multiple pluripotent stem cell types have been shown to exist in human, similar to the mouse

model, it will be important to characterize those stem cell types in detail and to analyze their

differences. We hypothesize that the various culture and derivation conditions will have a

substantial impact on the genomic stability and differentiation potential of the resulting stem cell

types.

Firstly, we will perform an in depth transcriptomic and (epi)genomic characterization and

comparison of our in-house established primed, naïve and IWP2-induced hESC lines. Secondly, by

performing single cell genomics, genomic heterogeneity within the different hESC cultures will be

revealed for the first time (WP1).

Next, we want to investigate the role of replicative stress as a driving force for genomic instability

in hESCs. For this, we will experimentally challenge our in-house derived primed, naïve and IWP2-

induced hESCs and monitor markers for replicative stress as well as their consequences on the

transcriptome and epigenome at different time points (WP2).

Finally, we want to compare the spontaneous, directed and targeted (neuronal and primordial

germ cell progenitor cells) differentiation potential of the different types of hESCs (WP3), which

will give important information for future possible regenerative applications.