title: The embryonic stem cell test as tool to assess structure-dependent teratogenicity: the case of valproic acid.
abstract: Teratogenicity can be predicted in vitro using the embryonic stem cell test (EST). The EST, which is based on the morphometric measurement of cardiomyocyte differentiation and cytotoxicity parameters, represents a scientifically validated method for the detection and classification of chemicals according to their teratogenic potency. Furthermore, an abbreviated protocol applying flow cytometry of intracellular marker proteins to determine differentiation into the cardiomyocyte lineage is available. Although valproic acid (VPA) is in worldwide clinical use as antiepileptic drug, it exhibits two severe side effects, i.e., teratogenicity and hepatotoxicity. These limitations have led to extensive research into derivatives of VPA. Here we chose VPA as model compound to test the applicability domain and to further evaluate the reliability of the EST. To this end, we study six closely related congeners of VPA and demonstrate that both the standard and the molecular flow cytometry-based EST are well suited to indicate differences in the teratogenic potency among VPA analogs that differ only in chirality or side chain length. Our data show that identical results can be obtained by using the standard EST or a shortened protocol based on flow cytometry of intracellular marker proteins. Both in vitro protocols enable to reliably determine differentiation of murine stem cells toward the cardiomyocyte lineage and to assess its chemical-mediated inhibition.
authors.affiliation: German Federal Institute for Risk Assessment (BfR), Center for Alternative Methods to Animal Experiments-ZEBET, 12277 Berlin, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Riebeling
authors.forename: Ralph
authors.initials: R
authors.lastname: Pirow
authors.forename: Klaus
authors.initials: K
authors.lastname: Becker
authors.forename: Roland
authors.initials: R
authors.lastname: Buesen
authors.forename: Daniel
authors.initials: D
authors.lastname: Eikel
authors.forename: Johanna
authors.initials: J
authors.lastname: Kaltenhäuser
authors.forename: Frauke
authors.initials: F
authors.lastname: Meyer
authors.forename: Heinz
authors.initials: H
authors.lastname: Nau
authors.forename: Birgitta
authors.initials: B
authors.lastname: Slawik
authors.forename: Anke
authors.initials: A
authors.lastname: Visan
authors.forename: Jutta
authors.initials: J
authors.lastname: Volland
authors.forename: Horst
authors.initials: H
authors.lastname: Spielmann
authors.forename: Andreas
authors.initials: A
authors.lastname: Luch
authors.forename: Andrea
authors.initials: A
authors.lastname: Seiler
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D055808:Drug Discovery; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects*; D005434:Flow Cytometry; D051379:Mice; D015394:Molecular Structure; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects*; D013329:Structure-Activity Relationship; D013723:Teratogens / Q000737:chemistry / Q000633:toxicity*; D018675:Toxicity Tests / Q000379:methods*; D014635:Valproic Acid / Q000031:analogs & derivatives / Q000737:chemistry / Q000633:toxicity*
chemical_list: D013723:Teratogens; D014635:Valproic Acid