Abstract
We investigated the gap junctional properties of human embryonic stem cells (hESC) cultivated in a serum-free system using sphingosine-1-phosphate and platelet-derived growth factor (S1P/PDGF). We compared this condition to hESC grown on Matrigel in mouse embryonic fibroblast conditioned medium (MEF-CM) or unconditioned medium (UM). We show that in all culture systems, hESC express connexins 43 and 45. hESC maintained in S1P/PDGF conditions and hESC grown in presence of MEF-CM are coupled through gap junctions while hESC maintained on Matrigel in UM do not exhibit gap junctional intercellular communication. In this latter condition, coupling was retrieved by addition of noggin, suggesting that BMP-like activity in UM inhibits gap junctional communication. Last, our data indicate that the closure of gap junctions by the decoupling agent alpha-glycyrrhetinic acid increases cell apoptosis and inhibits hESC colony growth. Altogether, these results suggest that gap junctions play an important role in hESC maintenance.
Publication types
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Comparative Study
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Apoptosis* / drug effects
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Bone Morphogenetic Proteins / metabolism
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Carrier Proteins / pharmacology
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Cell Communication / drug effects
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Cell Communication / physiology*
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Cell Culture Techniques
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Collagen / pharmacology
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Connexin 43 / metabolism
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Connexins / metabolism
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Culture Media, Conditioned
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Drug Combinations
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Embryo, Mammalian / cytology
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Gap Junctions / drug effects
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Gap Junctions / physiology*
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Glycyrrhetinic Acid / pharmacology
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Humans
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Laminin / pharmacology
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Lysophospholipids / pharmacology*
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Mice
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Platelet-Derived Growth Factor / pharmacology*
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Proteoglycans / pharmacology
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Sphingosine / analogs & derivatives*
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Sphingosine / pharmacology
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Stem Cells / cytology
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Stem Cells / drug effects*
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Stem Cells / physiology
Substances
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Bone Morphogenetic Proteins
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Carrier Proteins
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Connexin 43
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Connexins
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Culture Media, Conditioned
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Drug Combinations
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Laminin
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Lysophospholipids
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Platelet-Derived Growth Factor
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Proteoglycans
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connexin 45
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matrigel
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noggin protein
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sphingosine 1-phosphate
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Collagen
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Sphingosine
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Glycyrrhetinic Acid