Glycomic Signature of Mouse Embryonic Stem Cells During Differentiation*Corresponding Author(s): Rania Harfouche, BWH-HST Center for Biomedical Engineering, Harvard Medical School, 65 Landsdowne Street, Cambridge MA 02139, USA, Email: [email protected] Shiladitya Sengupta, BWH-HST Center for Biomedical Engineering, Harvard Medical School, 65 Landsdowne Street Cambridge MA 02139, USA
Received Date: Jul 22, 2013 / Accepted Date: Aug 21, 2013 / Published Date: Aug 24, 2013
Citation: Harfouche R, Ray S, Sanchez M, Dadwal U, Head SR, et al. (2013) Glycomic Signature of Mouse Embryonic Stem Cells During Differentiation. Cell Dev Biol 2:120.DOI: 10.4172/2168-9296.1000120
Copyright: © 2013 . This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Background: The glycome has emerged as a key regulator of cell fate, partly through its ability to potentiate the action of numerous signaling pathways. We recently demonstrated that a sulfated component of the glycome plays a critical role in promoting the differentiation of embryonic stem cell (ESC)-derived embryoid bodies by modulating downstream growth factors, such as the insulin-like growth factor (IGF) signaling axis. However, the exact components of the glycome which promote ESC differentiation versus stemness remain uncharacterized, due to the lack of a rapid, simple and easily quantifiable methodology. As a proof-of-concept in this study, we utilized a custom-made glycoarray in combination with bioinformatics and molecular biology tools in order to uncover novel glyco-signatures underlying ESC differention in an embryoid body model. A better elucidation of the glycomic transcriptomal signature underlying ESC differentiation would allow us to better manipulate these cells towards a desired lineage.
Method: We used a custom-designed Affymetrix microarray, the Glycogene-chip, to screen the transcriptome of differentiating embryoid bodies versus that of undifferentiated ESC. In conjunction with gene ontology, pathway analyses, real-time PCR and immunoblotting, we validated the involvement of the IGF family, and furthermore, uncovered novel differentially regulated genes belonging to the glycoprotein (Angiopoietin-1 and Angiopoietin-like members), sulfotransferase, sulfatase and glycosyltransferase families.
Conclusion: These results suggest that the Glycogene-chip, in conjunction with the embryoid body model, provides a fast and reliable tool to uncover novel glycomic signatures that are critical to maitain ESC stemness versus differentiation. In turn, this will allow us to understand the mechanisms governing ESC fate, bringing us one step closer towards finding a new paradigm for the regenerative medicine field.