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Imprint
Improvement of an in vitro stem cell assay for developmental toxicity: Establishing molecular endpoints of tissue specific development
Project
Project code: BfR-ZEBET-02-1334-142
Contract period: 01.09.2000
- 31.03.2004
Purpose of research: Applied research
The aim of this study is to improve an in vitro approach for developmental toxicity that is based on blastocyst-derived pluripotent embryonic stem (ES) cells of the mouse (cell line D3), the so-called embryonic stem cell test (EST). The EST represents a reliable, scientifically validated in vitro system for the classification of compounds according to their teratogenic potential based on the morphological analysis of beating cardiomyocytes in embryoid body (EB) outgrowths compared to cytotoxic effects on undifferentiated murine ES cells and differentiated 3T3 fibroblasts. In a joint project with major German pharmaceutical companies we attempt to expand the EST protocol by establishing molecular endpoints of differentiation (e.g. myocardial, neuronal, chondrogenic and osteogenic) in cultured ES cells. These improvements will help to identify more objective endpoints of differentiation other than the microscopic evaluation of beating cardiomyocytes and to adapt the EST to applications in high-throughput screening systems. We have studies the expression of cell type specific proteins in ES cell cultures in the presence of embryotoxic chemicals by intracellular flow cytometry. Our partners are focusing on endogenous gene expression in early development using real time PCR methods or the DNA microarray technique.
Section overview
Subjects
- Biotechnology
Framework programme
Funding programme
Excutive institution
BfR - Centre for Documentation and Evaluation of Alternatives to Animal Experiments (BfR - ZEBET)
