Self-renewal of human embryonic stem cells requires insulin-like growth factor-1 receptor and ERBB2 receptor signaling

Despite progress in developing defined conditions for human embryonic stem cell (hESC) cultures, little is known about the cell-surface receptors that are activated under conditions supportive of hESC self-renewal. A simultaneous interrogation of 42 receptor tyrosine kinases (RTKs) in hESCs followin...

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Bibliographic Details
Published in:Blood 2007-12, Vol.110 (12), p.4111-4119
Main Authors: Wang, Linlin, Schulz, Thomas C., Sherrer, Eric S., Dauphin, Derek S., Shin, Soojung, Nelson, Angelique M., Ware, Carol B., Zhan, Mei, Song, Chao-Zhong, Chen, Xiaoji, Brimble, Sandii N., McLean, Amanda, Galeano, Maria J., Uhl, Elizabeth W., D'Amour, Kevin A., Chesnut, Jonathan D., Rao, Mahendra S., Blau, C. Anthony, Robins, Allan J.
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal - pharmacology
Apoptosis - drug effects
Apoptosis - physiology
Benzothiazoles - pharmacology
Biological and medical sciences
Cell Differentiation - drug effects
Cell Differentiation - physiology
Cell Line
Cell Proliferation - drug effects
Culture Media, Conditioned
Embryonic Stem Cells - cytology
Embryonic Stem Cells - metabolism
Fibroblast Growth Factor 2 - pharmacology
Fibroblasts - cytology
Fibroblasts - metabolism
Hematologic and hematopoietic diseases
Humans
Medical sciences
Mice
Neuregulin-1 - pharmacology
Phosphorylation - drug effects
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - metabolism
Receptor, ErbB-2 - antagonists & inhibitors
Receptor, ErbB-2 - metabolism
Receptor, ErbB-3 - antagonists & inhibitors
Receptor, ErbB-3 - metabolism
Receptor, IGF Type 2 - antagonists & inhibitors
Receptor, IGF Type 2 - metabolism
Receptor, Insulin - antagonists & inhibitors
Receptor, Insulin - metabolism
Signal Transduction - drug effects
Signal Transduction - physiology
Stem Cells in Hematology
Tyrphostins - pharmacology
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Summary:Despite progress in developing defined conditions for human embryonic stem cell (hESC) cultures, little is known about the cell-surface receptors that are activated under conditions supportive of hESC self-renewal. A simultaneous interrogation of 42 receptor tyrosine kinases (RTKs) in hESCs following stimulation with mouse embryonic fibroblast (MEF) conditioned medium (CM) revealed rapid and prominent tyrosine phosphorylation of insulin receptor (IR) and insulin-like growth factor-1 receptor (IGF1R); less prominent tyrosine phosphorylation of epidermal growth factor receptor (EGFR) family members, including ERBB2 and ERBB3; and trace phosphorylation of fibroblast growth factor receptors. Intense IGF1R and IR phosphorylation occurred in the absence of MEF conditioning (NCM) and was attributable to high concentrations of insulin in the proprietary KnockOut Serum Replacer (KSR). Inhibition of IGF1R using a blocking antibody or lentivirus-delivered shRNA reduced hESC self-renewal and promoted differentiation, while disruption of ERBB2 signaling with the selective inhibitor AG825 severely inhibited hESC proliferation and promoted apoptosis. A simple defined medium containing an IGF1 analog, heregulin-1β (a ligand for ERBB2/ERBB3), fibroblast growth factor-2 (FGF2), and activin A supported long-term growth of multiple hESC lines. These studies identify previously unappreciated RTKs that support hESC proliferation and self-renewal, and provide a rationally designed medium for the growth and maintenance of pluripotent hESCs.
ISSN:0006-4971
1528-0020
DOI:10.1182/blood-2007-03-082586