Project description:The generation of specific types of neurons from stem cells offers important opportunities in regenerative medicine. However, future applications and proper verification of cell identities will require stringent ways to generate homogenous neuronal cultures. Here we show that under permissive culturing conditions individual transcription factors can induce a desired neuronal lineage from virtually all expressing cells by a mechanism resembling developmental binary cell fate switching. Such efficient selection of cell fate resulted in remarkable cellular enrichment that enabled global gene expression validation of generated neurons and identification of novel features in the studied cell lineages. Several sources of stem cells have a limited competence to differentiate into e.g. dopamine neurons. However, we show that the combination of factors that normally promote either regional or dedicated neuronal specification can overcome limitations in cellular competence and promote efficient reprogramming also in more remote neural contexts, including human neural progenitor cells. We used microarray analysis to verify the identity of several mESC derived neuronal cell types. By genome-wide gene expression comparisons we gained novel insights into molecular properties of several clinical relevant neuronal cell types. Total RNA was extracted from wild-type and transcription-factor induced mESC derived post-mitotic neurons and hybridized on Affymetrix arrays. In total triplicates of 9 different samples were analyzed. Wild-type samples served as control samples.
Project description:scRNA-seq of mouse embryonic stem cells (mESC) derived from four different genetic backgrounds grown in ground state conditions and differentiated towards an epiblast stem cell like (EpiSCL) population.
