Characterizing Changes in the Transcriptional Networks underlying Pluripotency in Mouse Embryonic Stem Cells upon the Induction of Differentiation

• Main Author: Schwartz, Michael Louis
• Other Authors: Mitchell, Jennifer
• Language: en_ca
• Published: 2012
• Subjects: Gene Expression; Pluripotency; Mouse Embryonic Stem Cells; Nanog; 0369
• Online Access: http://hdl.handle.net/1807/33522

Mouse embryonic stem cells (mESCs) are pluripotent cells capable of differentiating into all three germ layers present in the adult mouse. In this thesis, I have investigated the transcriptional changes that mESCs undergo as they are induced to differentiate towards the mesoderm lineage by 2i/LIF withdrawal and dimethyl sulfoxide (DMSO) treatment. 5 days of differentiation causes significant drops in expression of Sox2 and Oct4 primary transcript, while expression of Nanog and Kit significantly drops after only 1 day. It was determined that DMSO has no effect on the short-term changes in Nanog and Kit expression induced by 2i/LIF withdrawal. An expanded look at pluripotency-associated genes shows significant up-regulation of Oct4 and down-regulation of Klf4 and Stat3 following only 6 hours of 2i/LIF withdrawal. This data indicates that while some aspects of the transcriptional networks underlying pluripotency respond quickly to mesodermal differentiation cues, others remain unchanged for up to 5 days.