A stem cell-based reporter assay was developed to detect drug-induced alterations in the canonical Wnt/ß-catenin signaling pathway, which is involved in the regulation of early embryonic development. The so-called ReProGlo assay allows simultaneous determination of cell viability and luciferase reporter activity in a high throughput 96-well microtiter format. A clone of mouse embryonic stem (mES) cells stably expressing the SuperTopFlash reporter was established. This allows Wnt pathway activity determinations in undifferentiated mES cells and their differentiated descendants. Several test chemicals were analyzed in the new assay system. Known embryotoxicants like retinoic acid or lithium chloride induced concentration-dependent increases in reporter activity. The potency of yalproic acid and a series of structural analogs to activate the Wnt pathway correlated well with their reported teratogenic activity in the mouse. Cyclophosphamide was also active but only after metabolic activation by hepatocytes. The new test may help to predict embryotoxic potential of chemicals.
BREMER Susanne;
UIBEL Frederik;
STUMMANN Tina;
SCHWARZ Michael;
WEIMER Marc;
MUEHLEISEN Annette;
KÖHLE Christoph;
2010-10-19
PERGAMON-ELSEVIER SCIENCE LTD
JRC55844
0890-6238,
https://publications.jrc.ec.europa.eu/repository/handle/JRC55844,
10.1016/j.reprotox.2009.12.002,
This document is only visible at the Commission level.
You are not authorized to publish or distribute it outside the European Commission.
This is a public document. You can share this publication.
Additional supporting files
| File name | Description | File type | |
|---|
