Tracking mesoderm induction and its specification to the hemangioblast during embryonic stem cell differentiation

Fehling, Hans Jörg; Lacaud, Georges; Kubo, Atsushi; Kennedy, Marion; Robertson, Scott; Keller, Gordon; Kouskoff, Valérie

doi:10.1242/dev.00589

Cited by 462 publications

(576 citation statements)

References 52 publications

Supporting

Mentioning

534

Contrasting

Unclassified

Order By: Relevance

“…We observed mRNA within 2-3 days for nestin (an early neural ectoderm marker) and brachyury (Brachy, an early mesoderm marker), whereas β-H1 globin (β-globin, a mesoderm marker) and α-fetoprotein (α-FP, an endoderm marker) mRNAs accumulated later (days 5-6). These patterns of expression are as previously reported for multilineage differentiation following LIF withdrawal [8,9,32,[38][39][40][41].…”

Section: Progesterone Receptor Gene Is Expressed Early During Differesupporting

confidence: 87%

Differentiation of murine embryonic stem cells induces progesterone receptor gene expression

Sauter

McDermid

Weinberg

et al. 2005

Experimental Cell Research

View full text Add to dashboard Cite

The role of steroid hormone receptors in very early embryonic development remains unknown. Clearly, expression during organogenesis is important for tissue-specific development. However, progesterone receptor (PR) and estrogen receptors (ERα, ERβ), are expressed during early development through the blastocyst stage in mice and other species, and yet are not essential for embryonic viability. We have utilized the mouse embryonic stem (mES) cell model to investigate the regulated expression of these receptors during differentiation. Surprisingly, one of the earliest changes in gene expression in response to a differentiation signal observed is PR gene induction. It parallels the time course of expression for the patterning genes Hoxb1 and Hoxa5. Unexpectedly, PR gene expression is not regulated in an estrogen dependent manner by endogenous ERs or by transiently overexpressed ERα. Our results suggest a potentially novel mechanism of PR gene regulation within mES cells compared to adult tissues and the possibility of unique targets of PR action during early mES cell differentiation

show abstract

Section: Progesterone Receptor Gene Is Expressed Early During Differesupporting

confidence: 87%

Differentiation of murine embryonic stem cells induces progesterone receptor gene expression

Sauter

McDermid

Weinberg

et al. 2005

Experimental Cell Research

View full text Add to dashboard Cite

show abstract

“…Using an ES-cell line with the GFP cDNA targeted to the brachyury locus [43], we monitored early mesoderm development in differentiating cells by flow cytometry. Consistent with the described pattern of brachyury expression [43], GFP þ cells were detected between days 2 and 5 of differentiation in high glucose, with the greatest number of GFP þ cells occurring at days 3 and 4 ( Fig. 6A, 6B).…”

Section: Resultsmentioning

confidence: 99%

Mitochondrial Reactive Oxygen Species Mediate Cardiomyocyte Formation from Embryonic Stem Cells in High Glucose

et al. 2010

View full text Add to dashboard Cite

Accumulating evidence points to reactive oxygen species (ROS) as important signaling molecules for cardiomyocyte differentiation in embryonic stem (ES) cells. Given that ES cells are normally maintained and differentiated in medium containing supraphysiological levels of glucose (25 mM), a condition which is known to result in enhanced cellular ROS formation, we questioned whether this high glucose concentration was necessary for cardiomyocyte lineage potential. We show here that ES cells cultured in physiological glucose (5 mM), maintained their general stemness qualities but displayed an altered mitochondrial metabolism, which resulted in decreased ROS production. Furthermore, ES and induced pluripotent stem (iPS) cells differentiated in lower glucose concentrations failed to generate cardiomyocyte structures; an effect mimicked with antioxidant treatments using catalase, N-acetyl cysteine and mitoubiquinone, under high glucose conditions in ES cells. Molecular analysis revealed that ES cells differentiated in 5 mM glucose had reduced expression of the pro-cardiac NOX4 gene and diminished phosphorylation of p38 mitogen-activated protein kinase (MAPK), together with specific changes in the cardiac transcriptional network. These outcomes could be reversed by supplementation of low glucose cultures with ascorbic acid, paradoxically acting as a pro-oxidant. Furthermore, forced expression of an upstream p38 MAPK kinase (MKK6) could bypass the requirement for ROS during differentiation to cardiomyocytes under low glucose conditions, illustrating a key role for p38 in the cardiac differentiation program. Together these data demonstrate that endogenous ROS control is important for cardiomyocyte formation from ES cells, and furthermore that supraphysiological glucose, by supplying ROS, is absolutely required.

show abstract

“…We found that different lines could be distinguished by the expression of primitive streak or mesendoderm-associated genes. The Brachyury gene is frequently used as a marker to track early cellular differentiation into the mesodermal lineage in vitro [18,19]. However, the very early (E5.5) pregastrulation epiblast is negative for Brachyury expression.…”

Section: Discussionmentioning

confidence: 99%

Distinct Developmental Ground States of Epiblast Stem Cell Lines Determine Different Pluripotency Features

et al. 2011

View full text Add to dashboard Cite

Epiblast stem cells (EpiSCs) are pluripotent stem cells derived from mouse postimplantation embryos at embryonic day (E) 5.5-E7.5 at the onset of gastrulation, which makes them a valuable tool for studying mammalian postimplantation development in vitro. EpiSCs can also be reprogrammed into a mouse embryonic stem cell (mESC)-like state. Some reports have shown that the reversion of EpiSCs requires transcription factor overexpression, whereas others have suggested that use of stringent mESC culture conditions alone is sufficient for the reversion of EpiSCs. To clarify these discrepancies, we systematically compared a panel of independent EpiSC lines. We found that-regardless of the embryonic day of derivation-the different EpiSC lines shared a number of defining characteristics such as the ability to form teratomas. However, despite use of standard EpiSC culture conditions, some lines exhibited elevated expression of genes associated with mesendodermal differentiation. Pluripotency (Oct4) and mesodermal (Brachyury) marker genes were coexpressed in this subset of lines. Interestingly, the expression of mesendodermal marker genes was negatively correlated with the cells' ability to efficiently undergo neural induction. Moreover, these mesodermal marker gene-expressing cell lines could not be efficiently reverted to an mESC-like state by using stringent mESC culture conditions. Conversely, Brachyury overexpression diminished the reversion efficiency in otherwise Brachyury-negative lines. Overall, our data suggest that different EpiSC lines may undergo self-renewal into distinct developmental states, a finding with important implications for functional readouts such as reversion of EpiSCs to an mESC-like state as well as directed differentiation. STEM CELLS

show abstract

Tracking mesoderm induction and its specification to the hemangioblast during embryonic stem cell differentiation

Cited by 462 publications

References 52 publications

Differentiation of murine embryonic stem cells induces progesterone receptor gene expression

Differentiation of murine embryonic stem cells induces progesterone receptor gene expression

Mitochondrial Reactive Oxygen Species Mediate Cardiomyocyte Formation from Embryonic Stem Cells in High Glucose

Distinct Developmental Ground States of Epiblast Stem Cell Lines Determine Different Pluripotency Features

Contact Info

Product

Resources

About