Mapping the first stages of mesoderm commitment during differentiation of human embryonic stem cells
- Denis Evseenkoa,
- Yuhua Zhua,
- Katja Schenke-Laylandb,
- Jeffrey Kuoa,
- Brooke Latoura,
- Shundi Gea,
- Jessica Scholesa,
- Gautam Dravida,
- Xinmin Lia,
- W. Robb MacLellanb, and
- Gay M. Crooksa,1
- aDepartment of Pathology and Laboratory Medicine, and Broad Stem Cell Research Center and
- bCardiovascular Research Laboratory, David Geffen School of Medicine, University of California, Los Angeles, CA 90095
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Edited* by Owen N. Witte, Howard Hughes Medical Institute, UCLA, Los Angeles, CA, and approved June 25, 2010 (received for review February 19, 2010)
Abstract
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.
Footnotes
- 1To whom correspondence should be addressed. E-mail: gcrooks{at}mednet.ucla.edu.
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Author contributions: D.E. and G.M.C. designed research; D.E., Y.Z., K.S.-L., J.K., B.L., and S.G. performed research; K.S.-L., J.S., and X.L. contributed new reagents/analytic tools; D.E., K.S.-L., G.D., X.L., W.R.M., and G.M.C. analyzed data; and D.E. and G.M.C. wrote the paper.
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The authors declare no conflict of interest.
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↵*This Direct Submission article had a prearranged editor.
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This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1002077107/-/DCSupplemental.
