The role of Coup-TFII in the acquisition of cellular fates during the differentiation of embryonic stem cells

Abstract

Embryonic Stem Cells (ESC) are very important in researches that are looking for a complete understanding of the primordial embryonic development, in the development of cellular lineages and in the somatic cell reprogramming utilized in cellular therapy for tissue regeneration in many diseases. These cells have the capability of auto-renew and pluripotency, and cellular differentiation assays in vitro using ESC have been broadly use to understand the balance between undifferentiated and pluripotent states of these cells with the determination of cellular fate in vitro. The transcription factor COUP-TFII (Chicken Ovalbumin Promoter-transcription Factor II) has a fundamental role in the regulation of embryonic development and in the acquisition of specific fates, further it's involved in a complex regulatory circuitry that controls the pluriopotency of ESC. Order to analyse the possible role of COUP-TFII in the loss of pluripotency and the acquisition of specific cellular fates, this study intends to overexpress and silence the functions of this gene in Embryoid Bodies (EB) derived from ESC E14-Tg2a, followed by a characterization of molecular and morphological patterns of the cellular differentiation. For this, EBs with normal, increased and decreased expressions will be evaluated for pluripotency factors expression and markers of the three germ layers by immunohistochemistry markings and cellular quantifications, by flow cytometry, and the transcription, by qPCR. (AU)