Article

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

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.

Download full-text

Full-text

Available from: Katja Schenke-Layland
  • Source
    • "Primitive mesoderm (PMESO) has been isolated using the cell surface marker ROR2 (Drukker et al., 2012) and cardiac mesoderm (CMESO) is known to express PDGFR (Mummery et al., 2012). However, most candidate markers are, to variable extents, expressed on alternative cell types, and distinct lineages can require multiple surface markers for isolation (Evseenko et al., 2010). To date, no studies have globally assessed pluripotent-to-CM commitment using purified mesoderm cells. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The directed differentiation of human cardiomyocytes (CMs) from pluripotent cells provides an invaluable model for understanding mechanisms of cell fate determination and offers considerable promise in cardiac regenerative medicine. Here, we utilize a human embryonic stem cell suspension bank, produced according to a good manufacturing practice, to generate CMs using a fully defined and small molecule-based differentiation strategy. Primitive and cardiac mesoderm purification was used to remove non-committing and multi-lineage populations and this significantly aided the identification of key transcription factors, lncRNAs, and essential signaling pathways that define cardiomyogenesis. Global methylation profiles reflect CM development and we report on CM exon DNA methylation “memories” persisting beyond transcription repression and marking the expression history of numerous developmentally regulated genes, especially transcription factors.
    Full-text · Article · Jan 2016 · EBioMedicine
  • Source
    • "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). "
    [Show abstract] [Hide abstract]
    ABSTRACT: The latest discoveries and advanced knowledge in the fields of stem cell biology and developmental cardiology hold great promise for cardiac regenerative medicine, enabling researchers to design novel therapeutic tools and approaches to regenerate cardiac muscle for diseased hearts. However, progress in this arena has been hampered by a lack of reproducible and convincing evidence, which at best has yielded modest outcomes and is still far from clinical practice. To address current controversies and move cardiac regenerative therapeutics forward, it is crucial to gain a deeper understanding of the key cellular and molecular programs involved in human cardiogenesis and cardiac regeneration. In this review, we consider the fundamental principles that govern the "programming" and "reprogramming" of a human heart cell and discuss updated therapeutic strategies to regenerate a damaged heart. © 2015 The Authors.
    Full-text · Article · Feb 2015 · The EMBO Journal
  • Source
    • "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). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Distinguishing between perivascular cell types remains a hurdle in vascular biology due to overlapping marker expressions and similar functionalities. Clarifying and defining heterogeneities in vitro among perivascular cells could lead to improved cell-based tissue regeneration strategies and a better understanding of human developmental processes. We studied contractile vascular smooth muscle cells (vSMCs), synthetic vSMCs, and pericytes derived from a common human pluripotent stem cell source. Using in vitro cultures, we show unique cell morphology, subcellular organelle organization (namely endoplasmic reticulum, mitochondria, and stress fibers), and expression of smooth muscle myosin heavy chain and elastin for each cell type. While differences in extracellular matrix deposition and remodeling were less pronounced, the multipotency, in vivo, migratory, invasion, and contractile functionalities are distinctive for each cell type. Overall, we define a repertoire of functional phenotypes in vitro specific for each of the human perivascular cell types, enabling their study and use in basic and translational research.
    Full-text · Article · May 2014 · Stem Cell Reports
Show more