Abstract
BMP-11/GDF-11 and Myostatin/GDF-8 are both members of the TGF-beta superfamily that can activate SMAD2/3 phosphorylation via the type I receptors ALK4, ALK5, or ALK7. We tested the ability of BMP-11 and Myostatin to promote self-renewal of human embryonic stem cells (hESC) under feeder-free and serum-free culture conditions in short term (1 week) and medium term cultures (10 weeks). We show that hESC cultured in serum-free medium supplemented with either 20 ng/mL Myostatin or 20 ng/mL BMP-11 maintain the colony and cellular morphology of undifferentiated hESC, maintain POU5f1, NANOG, TRA-1-60, and SSEA4 expression, and display increased SMAD2/3 phosphorylation, similar to hESC cultured in mouse embryonic fibroblast feeder-CM or 20 ng/mL Activin-A. The type I TGF-beta receptor inhibitor SB431542 totally inhibited the maintenance activity of both Myostatin or BMP-11 supplemented medium. Our data show that members of the TGF-beta superfamily, other than Activin-A and GDF3, are able to maintain hES cells in an undifferentiated state under feeder free conditions.
MeSH terms
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Activin Receptors, Type I / antagonists & inhibitors
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Activin Receptors, Type I / biosynthesis
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Activins / pharmacology
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Animals
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Antigens, Surface / biosynthesis
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Benzamides / pharmacology
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Bone Morphogenetic Proteins / pharmacology*
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Cells, Cultured
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Coculture Techniques
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Dioxoles / pharmacology
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Embryonic Stem Cells / cytology*
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Embryonic Stem Cells / metabolism*
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Growth Differentiation Factors / pharmacology*
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Homeodomain Proteins / biosynthesis
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Humans
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Mice
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Myostatin / pharmacology*
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Nanog Homeobox Protein
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Octamer Transcription Factor-3 / biosynthesis
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Protein Serine-Threonine Kinases / antagonists & inhibitors
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Protein Serine-Threonine Kinases / biosynthesis
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Proteoglycans / biosynthesis
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Receptor, Transforming Growth Factor-beta Type I
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Receptors, Transforming Growth Factor beta / antagonists & inhibitors
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Receptors, Transforming Growth Factor beta / biosynthesis
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Smad2 Protein / metabolism
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Smad3 Protein / metabolism
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Stage-Specific Embryonic Antigens / biosynthesis
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Time Factors
Substances
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4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide
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Antigens, Surface
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Benzamides
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Bone Morphogenetic Proteins
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Dioxoles
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GDF11 protein, human
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Growth Differentiation Factors
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Homeodomain Proteins
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MSTN protein, human
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Myostatin
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NANOG protein, human
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Nanog Homeobox Protein
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Octamer Transcription Factor-3
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POU5F1 protein, human
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Proteoglycans
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Receptors, Transforming Growth Factor beta
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SMAD2 protein, human
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SMAD3 protein, human
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Smad2 Protein
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Smad3 Protein
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Stage-Specific Embryonic Antigens
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TRA-1-60 antigen, human
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activin A
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stage-specific embryonic antigen-4
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Activins
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Protein Serine-Threonine Kinases
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ACVR1B protein, human
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ACVR1C protein, human
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Activin Receptors, Type I
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Receptor, Transforming Growth Factor-beta Type I
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TGFBR1 protein, human
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Tgfbr1 protein, mouse