[Frontiers in Bioscience E4, 156-168, January 1, 2012]

[Frontiers in Bioscience E4, 156-168, January 1, 2012]

Effect of chemopreventive agents on differentiation of mouse embryonic stem cells

Vilas Wagh¹, Smita Jagtap¹, Kesavan Meganathan¹, Shiva Prasad Potta¹, Johannes Winkler¹, Juergen Hescheler¹, Agapios Sachinidis¹

¹Center of Physiology and Pathophysiology, Institute of Neurophysiology, Robert-Koch Str. 39, 50931 Cologne, Germany

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods: 3.1. Embryonic stem cells lines and culture; 3.2. Compounds; 3.3. Induction of ES cell Differentiation; 3.4. Measurement of growth inhibition of cells; 3.5. Measurement of ES cell differentiation inhibition; 3.6. RNA isolation and cDNA synthesis; 3.7. Semi-quantitative RT-PCR and quantitative RT-PCR; 3.8. Western blotting and Immunocytochemistry; 3.9. Statistics Analysis
4. Results: 4.1. Effects of EGCG, curcumin and quercetin on ESCs cytotoxicity; 4.2. Effects of the compounds on EB development; 4.3. Effects of the compounds on ESCs differentiation; 4.4. Effect of EGCG on the differentiation of ESCs into cardiomyocytes
5. Discussion
6. Acknowledgements
7. References

1. ABSTRACT

Chemopreventive agents are derived from edible plants and from ancient time is a part of daily intake for many humans and animals. There are several lines of compelling evidence from epidemiological, clinical and laboratory studies that these dietary constituents are associated in reducing cancer risks. However, developmental toxicity of these natural compounds cannot be excluded. In the present study, we examined the effect of chemopreventive agents on the differentiation of mouse embryonic stem cells (ESCs) as an in vitro embryotoxicity model. We assumed that inhibition of developmentally regulated genes in vitro might predict developmental toxicity also under in vivo conditions. We found that epigallocatechin gallate (EGCG) (20 然) induced the expression of mesodermal and cardiomyocyte genes and a significant increase in the number and the percentage of cardiomyocytes. The increase of the subpopulation correlated with higher numbers of beating foci and beating frequencies. Curcumin on the other hand at 0.4 mM was seen to enhance expression of ectodermal transcripts. Quercetin (2.5 然) was found to inhibit several developmentally regulated genes.