Proteomics Profiling of Human Embryonic Stem Cells in the Early Differentiation Stage

The regulatory pathways responsible for maintaining human embryonic stem cells (hESCs) in an undifferentiated state have yet to be elucidated. Since these pathways are thought to be governed by complex protein cues, deciphering the changes that occur in the proteomes of the ESCs during differentiati...

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Veröffentlicht in:Stem cell reviews 2012-03, Vol.8 (1), p.137-149
Hauptverfasser: Novak, Atara, Amit, Michal, Ziv, Tamar, Segev, Hanna, Fishman, Bettina, Admon, Arie, Itskovitz-Eldor, Joseph
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container_issue 1
container_start_page 137
container_title Stem cell reviews
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creator Novak, Atara
Amit, Michal
Ziv, Tamar
Segev, Hanna
Fishman, Bettina
Admon, Arie
Itskovitz-Eldor, Joseph
description The regulatory pathways responsible for maintaining human embryonic stem cells (hESCs) in an undifferentiated state have yet to be elucidated. Since these pathways are thought to be governed by complex protein cues, deciphering the changes that occur in the proteomes of the ESCs during differentiation is important for understanding the expansion and differentiation processes involved. In this study, we present the first quantitative comparison of the hESC protein profile in the undifferentiated and early differentiated states. We used iTRAQ (isobaric tags for relative and absolute quantification) labeling combined with two dimensional capillary chromatography coupled with tandem mass spectrometry (μLC-MS/MS) to achieve comparative proteomics of hESCs at the undifferentiated stage, and at 6, 48, and 72 h after initiation of differentiation. In addition, two dimensional electrophoresis (2-DE) was performed on differentiating hESCs at eleven points of time during the first 72 h of differentiation. The results indicate that during the first 48 h of hESC differentiation, many processes are initiated and are later reversed, including chromatin remodeling, heterochromatin spreading, a decrease in transcription and translation, a decrease in glycolytic proteins and cytoskeleton remodeling, and a decrease in focal and cell adhesion. Only 72 h after differentiation induction did the expression of the homeobox prox1 protein increase, indicating the beginning of developmental processes.
doi_str_mv 10.1007/s12015-011-9286-y
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Since these pathways are thought to be governed by complex protein cues, deciphering the changes that occur in the proteomes of the ESCs during differentiation is important for understanding the expansion and differentiation processes involved. In this study, we present the first quantitative comparison of the hESC protein profile in the undifferentiated and early differentiated states. We used iTRAQ (isobaric tags for relative and absolute quantification) labeling combined with two dimensional capillary chromatography coupled with tandem mass spectrometry (μLC-MS/MS) to achieve comparative proteomics of hESCs at the undifferentiated stage, and at 6, 48, and 72 h after initiation of differentiation. In addition, two dimensional electrophoresis (2-DE) was performed on differentiating hESCs at eleven points of time during the first 72 h of differentiation. The results indicate that during the first 48 h of hESC differentiation, many processes are initiated and are later reversed, including chromatin remodeling, heterochromatin spreading, a decrease in transcription and translation, a decrease in glycolytic proteins and cytoskeleton remodeling, and a decrease in focal and cell adhesion. 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subjects Biomedical and Life Sciences
Biomedical Engineering and Bioengineering
Cell adhesion
Cell Biology
Cell Differentiation
Cell Shape
Cells, Cultured
Chromatin remodeling
Chromatography
Cytoskeleton
Differentiation
Electrophoresis
Electrophoresis, Gel, Two-Dimensional
Embryo cells
Embryonic Stem Cells - metabolism
Embryonic Stem Cells - physiology
Gene Expression
Gene Expression Profiling
Gene Expression Regulation
Glycolysis
Heterochromatin
Homeobox
Humans
Karyotype
Life Sciences
Mass spectroscopy
Proteome - genetics
Proteome - isolation & purification
Proteome - metabolism
Proteomics
Regenerative Medicine/Tissue Engineering
Spreading
Stem Cells
Tandem Mass Spectrometry
Transcription
Translation
title Proteomics Profiling of Human Embryonic Stem Cells in the Early Differentiation Stage
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