Cytotoxicity test using blastocyst-derived euploid embryonal stem cells: a new approach to in vitro teratogenesis screening.
To develop a mammalian in vitro system for teratogenicity testing, cytotoxicity of xenobiotics was evaluated in pluripotent euploid embryonal stem cells (ESC) derived from mouse blastocysts. The dimethyl-thiazol-diphenyl tetrazolium bromide (MTT) assay was the most appropriate test system for cytotoxicity determinations with ESC. Only compounds that do not require metabolic activation were selected for testing from the database for validation of in vitro teratogenesis assays by Smith et al. Results obtained with ESC were compared to corresponding data from fibroblasts from day-14 mouse embryos to detect differences in sensitivity between undifferentiated and differentiated cells. ESC showed a higher sensitivity to known teratogens than fibroblast cultures, which allows calculation of a sensitivity ratio of "adult" cells (differentiated fibroblasts) to embryonal cells (undifferentiated ESC) in a mammalian system similar to the hydra assay. Although some xenobiotics had to be classified as false negatives in our system, the ESC cytotoxicity assay holds promise as a new in vitro screening assay in teratology.
Laschinski
G
G
ZEBET (Zentralstelle zur Erfassung und Bewertung von Ergänzungs- und Ersatzmethoden zum Tierversuch), Robert von Ostertag Institut, Berlin, Germany.
Vogel
R
R
Spielmann
H
H
D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents; D007202:Indicators and Reagents; D051379:Mice; D008815:Mice, Inbred Strains; D009499:Neutral Red; D009930:Organic Chemicals; D013234:Stem Cells / Q000187:drug effects*; D013723:Teratogens / Q000633:toxicity*; D013778:Tetrazolium Salts; D013844:Thiazoles
D004396:Coloring Agents; D007202:Indicators and Reagents; D009930:Organic Chemicals; D013723:Teratogens; D013778:Tetrazolium Salts; D013844:Thiazoles; C069245:kenacid blue; D009499:Neutral Red; C022616:thiazolyl blue