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.
Cloning and stem cells