Role of canonical Wnt/ β-catenin pathway in the commitment of embryonic stem cells during early differentiation

The different steps of embryonic formation in mammals have been deeply studied in the last few years. The possibility to isolate embryonic stem cells (ESCs) from embryos at the early stages of development has played a crucial role in identifying the mechanisms that drive the differentiation into embryonic and extraembryonic tissues. Different pathways are involved in the regulation of embryogenesis. It has been reported that Wnt/β-catenin pathway is crucial for the maintenance of stemness of ESCs, but little is know about its role in the differentiation of these cells. This research try to elucidate the molecular mechanisms of ESCs differentiation induced upon Wnt canonical pathway inhibition with different approaches: silencing of β-catenin (positive regulator of transcription), overexpression of TCF3 (negative regulator of transcription) and XAV939 (small drug inhibiting the upstream signalling). The differentiation process of ESCs has been monitored by checking the expression levels (RT-PCR) of markers of pluripotency, differentiation (to trophoblast, epiblast or primitive endoderm) also evaluating canonical Wnt target genes. In the next experiments we aim to confirm the role of the identified target genes in driving the process of differentiation that we observe. The knowledge of the canonical Wnt/β-catenin in the initial steps of embryo differentiation is essential also to shed light on diseases caused by defects in the embryonic development.status: publishe