An evaluation of the chick cardiomyocyte micromass system for identification of teratogens in a blind trial.
The chick micromass culture system has advantages over the validated rat system - ready availability and non-culling of the donor parent - but needs to give comparable results. This study confirmed comparability and the ability to extend the system to cover cardiac effects. It was also compared with the validated embryonic stem cell cardiomyocyte model. A teratogen and paired non-teratogen with known in vivo effects were used. Differential effects were measured via changes in cell protein content, cell viability (resazurin reduction and neutral red uptake), and cell contractility. Results showed that teratogens [L-ethionine, 5-fluorouracil and sulphisoxazole] could be distinguished from non-teratogens [DL-methionine, 6-methyluracil and sulphanilamide respectively]. Dichloroacetone and dichloropropanol affected embryonic stem cells but not the micromass; dichloropropanol had a greater effect than dichloroacetone. This approach revealed differential effects on contractility independent of effects on activity/viability, whilst the total cell protein remained unchanged. We suggest that pre-validation of this system should be examined.
Hurst
Helen
H
FRAME Alternatives Laboratories, School of Biomedical Sciences, University of Nottingham, Nottingham NG7 2UH, United Kingdom.
Clothier
Richard H
RH
Pratten
Margaret
M
D000096:Acetone / Q000031:analogs & derivatives / Q000633:toxicity; D000826:Animal Testing Alternatives; D000818:Animals; D002452:Cell Count; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002469:Cell Separation; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002642:Chick Embryo; D003470:Culture Media / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian; D053595:Embryonic Stem Cells / Q000187:drug effects; D005001:Ethionine / Q000633:toxicity; D005472:Fluorouracil / Q000633:toxicity; D007202:Indicators and Reagents / Q000378:metabolism; D008715:Methionine / Q000633:toxicity; D009200:Myocardial Contraction / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects; D009499:Neutral Red / Q000378:metabolism; D009930:Organic Chemicals / Q000378:metabolism; D010078:Oxazines / Q000032:analysis; D000077145:Sulfanilamide; D013424:Sulfanilamides / Q000633:toxicity; D013444:Sulfisoxazole / Q000633:toxicity; D013723:Teratogens / Q000633:toxicity*; D013997:Time Factors; D018675:Toxicity Tests / Q000379:methods*; D014498:Uracil / Q000031:analogs & derivatives / Q000633:toxicity; D014966:Xanthenes / Q000032:analysis; D000517:alpha-Chlorohydrin / Q000031:analogs & derivatives / Q000633:toxicity
D003470:Culture Media; D007202:Indicators and Reagents; D009930:Organic Chemicals; D010078:Oxazines; D013424:Sulfanilamides; D013723:Teratogens; D014966:Xanthenes; C033489:1,3-dichloro-2-propanol; C069245:kenacid blue; D000096:Acetone; C005843:resazurin; D000077145:Sulfanilamide; D009499:Neutral Red; D014498:Uracil; C008378:6-methyluracil; D013444:Sulfisoxazole; D000517:alpha-Chlorohydrin; D008715:Methionine; D005472:Fluorouracil; C047675:1,3-dichloroacetone; D005001:Ethionine