Abstract
We report that a single growth factor, NM23-H1, enables serial passaging of both human ES and iPS cells in the absence of feeder cells, their conditioned media or bFGF in a fully defined xeno-free media on a novel defined, xeno-free surface. Stem cells cultured in this system show a gene expression pattern indicative of a more "naïve" state than stem cells grown in bFGF-based media. NM23-H1 and MUC1* growth factor receptor cooperate to control stem cell self-replication. By manipulating the multimerization state of NM23-H1, we override the stem cell's inherent programming that turns off pluripotency and trick the cells into continuously replicating as pluripotent stem cells. Dimeric NM23-H1 binds to and dimerizes the extra cellular domain of the MUC1* transmembrane receptor which stimulates growth and promotes pluripotency. Inhibition of the NM23-H1/MUC1* interaction accelerates differentiation and causes a spike in miR-145 expression which signals a cell's exit from pluripotency.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Antibodies, Monoclonal / immunology
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Antibodies, Monoclonal / metabolism
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Binding, Competitive
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Biomarkers / metabolism
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Cell Differentiation
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Cells, Cultured
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Culture Media, Conditioned / pharmacology
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Embryonic Stem Cells / cytology
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Embryonic Stem Cells / drug effects
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Embryonic Stem Cells / metabolism
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Fibroblast Growth Factor 2 / pharmacology
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Fibroblasts / metabolism
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Humans
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Induced Pluripotent Stem Cells / cytology
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Induced Pluripotent Stem Cells / drug effects
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Induced Pluripotent Stem Cells / metabolism
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Ligands
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MicroRNAs / genetics
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MicroRNAs / metabolism
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Mucin-1 / immunology
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Mucin-1 / metabolism
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NM23 Nucleoside Diphosphate Kinases / chemistry
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NM23 Nucleoside Diphosphate Kinases / metabolism
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NM23 Nucleoside Diphosphate Kinases / pharmacology*
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Protein Binding / drug effects
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Protein Multimerization
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Stem Cells / cytology
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Stem Cells / drug effects*
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Stem Cells / metabolism
Substances
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Antibodies, Monoclonal
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Biomarkers
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Culture Media, Conditioned
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Ligands
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MIRN145 microRNA, human
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MicroRNAs
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Mucin-1
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NM23 Nucleoside Diphosphate Kinases
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Fibroblast Growth Factor 2
Grants and funding
The work of GL was supported by the National Science Foundation under Grant NSF-CDI ECC-0835847. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.