Stable Isotope Labelling with Amino Acids in Cell Culture for Human Embryonic Stem Cell Proteomic Analysis

2012 ◽  
pp. 297-305 ◽  
Author(s):  
Linda Harkness ◽  
Tatyana A. Prokhorova ◽  
Moustapha Kassem ◽  
Blagoy Blagoev
Keyword(s):  
Amino Acids ◽  
Cell Culture ◽  
Stem Cell ◽  
Stable Isotope ◽  
Embryonic Stem Cell ◽  
Proteomic Analysis ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Stable Isotope Labelling ◽  
Isotope Labelling

scholarly journals Dynamic Proteomic Analysis of Pancreatic Mesenchyme Reveals Novel Factors That Enhance Human Embryonic Stem Cell to Pancreatic Cell Differentiation

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Holger A. Russ ◽  
Limor Landsman ◽  
Christopher L. Moss ◽  
Roger Higdon ◽  
Renee L. Greer ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Cell Differentiation ◽  
Beta Cell ◽  
Embryonic Stem Cell ◽  
Proteomic Analysis ◽  
Human Embryonic Stem Cell ◽  
Pancreatic Beta Cell ◽  
Embryonic Stem ◽  
Pancreatic Cell

Current approaches in human embryonic stem cell (hESC) to pancreatic beta cell differentiation have largely been based on knowledge gained from developmental studies of the epithelial pancreas, while the potential roles of other supporting tissue compartments have not been fully explored. One such tissue is the pancreatic mesenchyme that supports epithelial organogenesis throughout embryogenesis. We hypothesized that detailed characterization of the pancreatic mesenchyme might result in the identification of novel factors not used in current differentiation protocols. Supplementing existing hESC differentiation conditions with such factors might create a more comprehensive simulation of normal development in cell culture. To validate our hypothesis, we took advantage of a novel transgenic mouse model to isolate the pancreatic mesenchyme at distinct embryonic and postnatal stages for subsequent proteomic analysis. Refined sample preparation and analysis conditions across four embryonic and prenatal time points resulted in the identification of 21,498 peptides with high-confidence mapping to 1,502 proteins. Expression analysis of pancreata confirmed the presence of three potentially important factors in cell differentiation: Galectin-1 (LGALS1), Neuroplastin (NPTN), and the Lamininα-2 subunit (LAMA2). Two of the three factors (LGALS1 and LAMA2) increased expression of pancreatic progenitor transcript levels in a published hESC to beta cell differentiation protocol. In addition, LAMA2 partially blocks cell culture induced beta cell dedifferentiation. Summarily, we provide evidence that proteomic analysis of supporting tissues such as the pancreatic mesenchyme allows for the identification of potentially important factors guiding hESC to pancreas differentiation.

scholarly journals Traditional Human Embryonic Stem Cell Culture

2011 ◽  
pp. 107-123 ◽  
Author(s):  
Philip H. Schwartz ◽  
David J. Brick ◽  
Hubert E. Nethercott ◽  
Alexander E. Stover
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Stem Cell Culture

Human Embryonic Stem Cell Culture

2007 ◽  
pp. 3-17 ◽  
Author(s):  
Rodolfo Gonzalez ◽  
Jeanne F. Loring ◽  
Robin L. esselschmidt ◽  
Philip H. Schartz
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Stem Cell Culture

scholarly journals Characterization of integrin engagement during defined human embryonic stem cell culture

2009 ◽  
Vol 24 (4) ◽  
pp. 1056-1065 ◽  
Author(s):  
Ying Meng ◽  
Shawdee Eshghi ◽  
Ying J. Li ◽  
Ray Schmidt ◽  
David V. Schaffer ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Stem Cell Culture

A quantitative proteomic analysis of lung epithelial (A549) cells infected with 2009 pandemic influenza A virus using stable isotope labelling with amino acids in cell culture

PROTEOMICS ◽  
2012 ◽  
Vol 12 (9) ◽  
pp. 1431-1436 ◽  
Author(s):  
Brian K. Dove ◽  
Rebecca Surtees ◽  
Thomas J.H. Bean ◽  
Diane Munday ◽  
Helen M. Wise ◽  
...  
Keyword(s):  
Amino Acids ◽  
Cell Culture ◽  
Stable Isotope ◽  
Influenza A Virus ◽  
Pandemic Influenza ◽  
Influenza A ◽  
A549 Cells ◽  
Stable Isotope Labelling ◽  
Isotope Labelling ◽  
Lung Epithelial
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