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Epicardial Outgrowth Culture Assay and Ex Vivo Assessment of Epicardial-derived Cell Migration
Published on: March 18, 2016
Wt1 and epicardial fate mapping.
Carsten Rudat1, Andreas Kispert
1Institut für Molekularbiologie, Medizinische Hochschule Hannover, Hannover, Germany.
Circulation Research
|June 14, 2012
Summary
Wt1-based lineage tracing systems are unreliable for mapping epicardial cell fate in mice due to Wt1 gene expression in endothelial cells and inconsistent recombination. These findings question previous claims of epicardium-derived cardiomyocytes.
Area of Science:
- Developmental Biology
- Cardiovascular Research
- Genetics
Background:
- The embryonic epicardium is a vital source for cardiac structures and coronary vessels.
- Previous studies suggested epicardium-derived cells contribute to myocardial development in mice using Wt1-based lineage tracing.
Purpose of the Study:
- To evaluate the suitability of Wt1-driven genetic tools for accurately tracing epicardial cell fate during mouse heart development.
Main Methods:
- Utilized in situ hybridization and immunofluorescence to detect Wt1 expression.
- Analyzed Wt1(creEGFP) and Wt1(creERT2) mouse lines for recombination efficiency and specificity.
- Examined reporter gene expression in various cardiac cell types.
Main Results:
- Endogenous Wt1 expression was found in epicardial and endothelial cells.
- Wt1(creEGFP) showed widespread, random recombination, including in early cardiomyocytes.
- Wt1(creERT2) exhibited inefficient and variable epicardial recombination, with no cardiomyocyte contribution observed.
Conclusions:
- Common Wt1-based lineage tracing methods are inadequate for epicardial fate mapping.
- Issues include Wt1 expression in non-epicardial cells and unreliable recombination.
- Current evidence does not support epicardial cells differentiating into cardiomyocytes in mice.
