Use of murine embryonic stem cells in embryotoxicity assays: the embryonic stem cell test

Seiler, A.; Buesen, R.; Visan, A.; Spielmann, H.

doi:10.1385/1-59745-037-5:371

Veröffentlicht

Use of murine embryonic stem cells in embryotoxicity assays: the embryonic stem cell test

Seiler, A.; Buesen, R.; Visan, A.; Spielmann, H.

The embryonic stem cell test (EST) takes advantage of the potential of murine embryonic stem (ES) cells to differentiate in culture to test embryotoxicity in vitro. The EST represents a 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 outgrowths compared to cytotoxic effects on murine ES cells and differentiated 3T3 fibroblasts. Through a number of prevalidation and validation studies, the EST has been demonstrated to be a reliable alternative method for embryotoxicity testing based on the most important mechanisms in embryotoxicity-cytotoxicity and differentiation--as well as on differences in sensitivity between differentiated and embryonic tissues. Improvements of the EST protocol using flow cytometry analysis showed that differential expression of sarcomeric myosin heavy chain and alpha-actinin proteins quantified under the influence of a test compound is a useful marker for detecting potential teratogenicity. The in vitro embryotoxicity test described in this chapter is rapid, simple, and sensitive and can be usefully employed as a component of the risk/hazard assessment process

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Erschienen in:: Methods in Molecular Biology
Vol. 329(2006), S. 371-395
Band:: 329
Datum der Veröffentlichung:: 2006
DOI:: 10.1385/1-59745-037-5:371
Sprache:: Englisch
Ressourcentyp:: Text
Schlagwörter:: 3T3 Cells; alternative; alternative methods; analysis; animal; ANIMAL-EXPERIMENTS; Animals; article; ASSAY; assessment; Berlin; BfR; CELL; Cell Culture Techniques; Cell Differentiation; Cell Line; CELLS; classification; compounds; CULTURE; cytology; DIFFERENCE; Documentation; drug effects; Drug Evaluation,Preclinical; effect; EFFECTS; Embryo; embryonic stem cell test; embryonic stem cells; embryotoxicity; EMBRYOTOXICITY TEST; evaluation; EXPRESSION; Fibroblasts; Flow Cytometry; Germany; In Vitro; IN-VITRO; MECHANISM; MECHANISMS; metabolism; method; methods; Mice; Myocytes,Cardiac; Myosin Heavy Chains; NUMBER; Pluripotent Stem Cells; potential; prevalidation; protein; Proteins; PROTOCOL; Risk; Risk Assessment; RISK-ASSESSMENT; SENSITIVITY; Solvents; Staining and Labeling; Stem Cells; study; SYSTEM; Teratogens; Testing; Tetrazolium Salts; Thiazoles; tissue; tissues; toxicity; validation; validation study; VITRO; ZEBET
DDC-Sachgruppe der DNB:: 600 Technik
Link URL:: http://link.springer.com/protocol/10.1385%2F1-59745-037-5%3A371
Einrichtung:: Bundesinstitut für Risikobewertung, Bundesinstitut für Risikobewertung (bis Juni 2014), Abteilung 3 - Wissenschaftliche Querschnittsaufgaben (bis Juni 2014)