Molecular markers in embryonic stem cells.
Embryonic stem cells are pluripotent cells derived from mouse blastocysts, which have the capacity to differentiate in vitro into a wide variety of cell types. Based on this potential the embryonic stem cell test (EST) has been developed, which represents an assay system for the classification of compounds for their teratogenic potential, based on the morphological evaluation of contracting myocard cells compared to the cytotoxic effects on undifferentiated stem cells and adult 3T3 fibroblasts. To expand the EST, the quantitative expression of the alpha- and beta-myosin heavy chain (MHC) genes under the influence of test compounds was studied employing real-time TaqMan PCR analysis. The molecular evaluation of the MHC genes allows a higher sensitivity for the classification of substances and the transfer of the EST to the molecular level allows to start experimental procedures at day 9 of culture. Thus, the modulated EST holds promise as a new easily quantifiable in vitro screening assay in teratology.
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Bayer AG, Research Toxicology, 42096 Wuppertal, Germany. nicole.melzer.nm.@bayer.ag.de
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Kempka
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Hildebrand
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Ahr
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D016475:3T3 Cells; D000826:Animal Testing Alternatives; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D003114:Colony-Forming Units Assay; D004305:Dose-Response Relationship, Drug; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D005819:Genetic Markers; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018995:Myosin Heavy Chains / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013723:Teratogens / Q000145:classification / Q000633:toxicity; D013778:Tetrazolium Salts / Q000378:metabolism; D024723:Ventricular Myosins / Q000235:genetics / Q000378:metabolism
D005819:Genetic Markers; D013723:Teratogens; D013778:Tetrazolium Salts; D024723:Ventricular Myosins; D018995:Myosin Heavy Chains