Author :
Release : 2014
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Kind : eBook
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Book Synopsis Engineering Density-dependent Signaling Pathways to Regulate Human Pluripotent Stem Cell Fate by :
Download or read book Engineering Density-dependent Signaling Pathways to Regulate Human Pluripotent Stem Cell Fate written by . This book was released on 2014. Available in PDF, EPUB and Kindle. Book excerpt: Much progress has been made in elucidating molecular pathways governing human pluripotent stem cell (hPSC) fate decisions and controlling differentiation fates to generate desired cell types. However, the transition from 2D monolayer culture to a more physiologically-relevant, 3D microenvironment brings various additional factors that affect stem cell fates, including soluble factors, extracellular matrix, mechanical forces and intercellular interactions. These mechanisms must be elucidated in order to completely realize the downstream potential of hPSCs. My research has utilized a 3D microwell array system to modulate the intercellular interactions of hPSCs and evaluate the effects of 3D cell interactions on developmental signaling pathways and differentiation. In addition to 3D culture, we also varied 2D culture densities to identify developmentally important signaling pathways that were modulated. To this end, we focused on the Hippo pathway due to its involvement in cell contact inhibition and hPSC pluripotency. We observed that at higher densities, YAP phosphorylation and localization to the cytoplasm increased, which led to decreased Hippo transcriptional activity, as measured using YAP responsive luciferase reporter cell lines. We also demonstrated that the density-dependent increase of neuroepithelial cell conversion rates from increasing density could be recapitulated by the knockdown of YAP. Taken together, these findings provide evidence that understanding the effects of specific 3D microenvironmental cues, such as cell density, in regulating hPSCs is relevant to developing physiologically-relevant cells and tissues.
