Mapping the first stages of mesoderm commitment during differentiation of human embryonic stem cells

Department of Pathology and Laboratory Medicine, and Broad Stem Cell Research Center and Cardiovascular Research Laboratory, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 08/2010; 107(31):13742-7. DOI: 10.1073/pnas.1002077107
Source: PubMed


Our understanding of how mesodermal tissue is formed has been limited by the absence of specific and reliable markers of early mesoderm commitment. We report that mesoderm commitment from human embryonic stem cells (hESCs) is initiated by epithelial-to-mesenchymal transition (EMT) as shown by gene expression profiling and by reciprocal changes in expression of the cell surface proteins, EpCAM/CD326 and NCAM/CD56. Molecular and functional assays reveal that the earliest CD326-CD56+ cells, generated from hESCs in the presence of activin A, BMP4, VEGF, and FGF2, represent a multipotent mesoderm-committed progenitor population. CD326-CD56+ progenitors are unique in their ability to generate all mesodermal lineages including hematopoietic, endothelial, mesenchymal (bone, cartilage, fat, fibroblast), smooth muscle, and cardiomyocytes, while lacking the pluripotency of hESCs. CD326-CD56+ cells are the precursors of previously reported, more lineage-restricted mesodermal progenitors. These findings present a unique approach to study how germ layer specification is regulated and offer a promising target for tissue engineering.

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    • "Fgf10 is also expressed in the SHF (Marguerie et al, 2006). In human ESCs, FGF2, in combination with Activin and BMP4, is known to specifically promote mesoderm-committed precursor formation (Evseenko et al, 2010). "
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    • "These data suggest the emergence of a mesodermal population. Furthermore, our day 6 differentiating cells were assessed for the expression of markers indicative of several intermediate lineages: neuroectoderm (nestin), lateral plate mesoderm (KDR), paraxial mesoderm (Pax1) (Cheung et al., 2012), early mesoderm (CD56) (Evseenko et al., 2010), and general mesoderm (CD73) (Boyd et al., 2009; Vodyanik et al., 2010). We found that a small fraction of day 6 differentiating cells was positive for nestin (Figure 1B). "
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    • "E-cadherin positivity gradually decreased during further differentiation , suggesting that the cells had undergone epithelial-to-mesenchymal (EMT) transition. EMT can be considered as an early marker of differentiation , as mesodermal commitment from hESCs was shown to be initiated by EMT (Evseenko et al., 2010). Parallel to the gradually loss of E-cadherin positivity, the proportion of PDGFRA positive cells within MIXL1-GFP + cells continuously increased, until the downregulation of MIXL1 occurred at about day 12. "
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