Dataset Information

Activin A and BMP4 Signaling for Efficient Cardiac Differentiation of H7 and H9 Human Embryonic Stem Cells.

ABSTRACT: The generation of cardiomyocytes from human embryonic stem cells (hESC) enables a variety of potential therapeutic and diagnostic applications. However, progress is challenged by the low efficiency of cardiomyocyte differentiation. Recently, Kattman et al., 2011 showed that individual hESC lines required proper balance of the Activin A and BMP4 signaling for efficient cardiac differentiation, presenting their differentiation protocols for several human and mouse ESC lines. However, two of the most utilized hESC lines, H7 and H9, were not included. Therefore, we set out to verify the published methodology for highly efficient cardiac specification and investigate the cardiac differentiation in the H7 and H9 ESC lines. Our studies examined a range of time points for the initial culture of hESC as embryoid bodies (EB) prior to transfer to monolayer culture, as well as, concentrations of Activin A and BMP4 in the medium formulations. The results highlight an efficient protocol for reproducibly generating cultures with approximately 50% cardiomyocytes from H7 and H9 ESC lines.

SUBMITTER: Sa S

PROVIDER: S-EPMC3908295 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Activin A and BMP4 Signaling for Efficient Cardiac Differentiation of H7 and H9 Human Embryonic Stem Cells.

Sa S S McCloskey Ke K

Journal of stem cells & regenerative medicine 20121126 3

The generation of cardiomyocytes from human embryonic stem cells (hESC) enables a variety of potential therapeutic and diagnostic applications. However, progress is challenged by the low efficiency of cardiomyocyte differentiation. Recently, Kattman et al., 2011 showed that individual hESC lines required proper balance of the Activin A and BMP4 signaling for efficient cardiac differentiation, presenting their differentiation protocols for several human and mouse ESC lines. However, two of the mo ...[more]

PMID: 24693198

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Project description:Previous studies show that both activin and Bmp4 act as crucial mesenchymal odontogenic signals during early tooth development. Remarkably, mice lacking activin-βA ( Inhba-/-) and mice with neural crest-specific inactivation of Bmp4 ( Bmp4ncko/ncko) both exhibit bud-stage developmental arrest of the mandibular molar tooth germs while their maxillary molar tooth germs completed morphogenesis. In this study, we found that, whereas expression of Inhba and Bmp4 in the developing tooth mesenchyme is independent of each other, Bmp4ncko/nckoInhba-/- compound mutant mice exhibit early developmental arrest of all tooth germs. Moreover, genetic inactivation of Osr2, a negative regulator of the odontogenic function of the Bmp4-Msx1 signaling pathway, rescues mandibular molar morphogenesis in Inhba-/- embryos. We recently reported that Osr2 and the Bmp4-Msx1 pathway control the bud-to-cap transition of tooth morphogenesis through antagonistic regulation of expression of secreted Wnt antagonists, including Dkk2 and Sfrp2, in the developing tooth mesenchyme. We show here that expression of Dkk2 messenger RNAs was significantly upregulated and expanded into the tooth bud mesenchyme in Inhba-/- embryos in comparison with wild-type littermates. Furthermore, in utero treatment with either lithium chloride, an agonist of canonical Wnt signaling, or the DKK inhibitor IIIC3a rescued mandibular molar tooth morphogenesis in Inhba-/- embryos. Together with our finding that the developing mandibular molar tooth bud mesenchyme expresses significantly higher levels of Dkk2 than the developing maxillary molar tooth mesenchyme, these data indicate that Bmp4 and activin signaling pathways converge on activation of the Wnt signaling pathway to promote tooth morphogenesis through the bud-to-cap transition and that the differential effects of loss of activin or Bmp4 signaling on maxillary and mandibular molar tooth morphogenesis are mainly due to the differential expression of Wnt antagonists, particularly Dkk2, in the maxillary and mandibular tooth mesenchyme.

Dataset Information

Activin A and BMP4 Signaling for Efficient Cardiac Differentiation of H7 and H9 Human Embryonic Stem Cells.

Publications

Activin A and BMP4 Signaling for Efficient Cardiac Differentiation of H7 and H9 Human Embryonic Stem Cells.

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