BMP-11 and myostatin support undifferentiated growth of human embryonic stem cells in feeder-free cultures.
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.
Hannan
Nicholas R F
NR
Department of Anatomy and Developmental Biology, Monash University, Clayton, Vic 3168, Australia.
Jamshidi
Pegah
P
Pera
Martin F
MF
Wolvetang
Ernst J
EJ
D030201:Activin Receptors, Type I / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D028341:Activins / Q000494:pharmacology; D000818:Animals; D000954:Antigens, Surface / Q000096:biosynthesis; D001549:Benzamides / Q000494:pharmacology; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002478:Cells, Cultured; D018920:Coculture Techniques; D004149:Dioxoles / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D055412:Growth Differentiation Factors / Q000494:pharmacology*; D018398:Homeodomain Proteins / Q000096:biosynthesis; D006801:Humans; D051379:Mice; D055435:Myostatin / Q000494:pharmacology*; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D011509:Proteoglycans / Q000096:biosynthesis; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D055746:Stage-Specific Embryonic Antigens / Q000096:biosynthesis; D013997:Time Factors
C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D000954:Antigens, Surface; D001549:Benzamides; D019485:Bone Morphogenetic Proteins; D004149:Dioxoles; C492266:GDF11 protein, human; D055412:Growth Differentiation Factors; D018398:Homeodomain Proteins; C525911:MSTN protein, human; D055435:Myostatin; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D011509:Proteoglycans; D018125:Receptors, Transforming Growth Factor beta; C497782:SMAD2 protein, human; C497790:SMAD3 protein, human; D051899:Smad2 Protein; D051900:Smad3 Protein; D055746:Stage-Specific Embryonic Antigens; C107064:TRA-1-60 antigen, human; C422405:activin A; C066584:stage-specific embryonic antigen-4; D028341:Activins; D017346:Protein Serine-Threonine Kinases; C498932:ACVR1B protein, human; C424205:ACVR1C protein, human; D030201:Activin Receptors, Type I; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625841:TGFBR1 protein, human; C000625842:Tgfbr1 protein, mouse