Automated analysis of contractility in the embryonic stem cell test, a novel approach to assess embryotoxicity.
The embryonic stem cell test (EST) is an ECVAM-validated assay to detect embryotoxicity. The output of the assay is the effect of test compounds on the differentiation of murine-derived embryonic stem cells (D3 cells), recorded by visual analysis of contracting cardiomyocyte-like cells. Incorporation of a system to assess the contractility in an automated manner is proposed, to increase the throughput in the EST independent of observer bias. The automated system is based on image recording of each well, resulting in the area (pixels) and frequency of contractility (Hz). Four test compounds were assessed for their embryotoxic potency in the 96-well version of the EST, with both manual and automated analysis: 6-Aminonicotinamide, Valproic Acid, Boric Acid, and Penicillin G. There was no statistically significant difference in the outcome of both methods in the fraction of contractility (p<0.05), resulting in the same rank-order of Relative Embryotoxic Potency (REP) values: 6-aminonicotinamide (1)>valproic acid (0.007-0.013)>Boric Acid (0.002-0.005)>Penicillin G (0.00001). The automated image recording of contractile cardiomyocyte-like cells in the EST allows for an unbiased high throughput method to assess the embryotoxic potency of test compounds, resulting in an outcome comparable to manual analysis.
Johnson&Johnson Pharmaceutical R&D (J&J-PRD), Department of Genetic and Exploratory Toxicology, Turnhoutseweg 30, B-2340 Beerse, Belgium. APeters5@prdbe.jnj.com
Annelieke K
AK
Peters
Gert Van de
GV
Wouwer
Barbara
B
Weyn
Geert R
GR
Verheyen
Philippe
P
Vanparys
Jacques Van
JV
Gompel
D000818:Animals; D001331:Automation; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D053595:Embryonic Stem Cells / Q000187:drug effects*; D051379:Mice; D009200:Myocardial Contraction / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism; D015588:Observer Variation; D013723:Teratogens / Q000633:toxicity*; D018675:Toxicity Tests / Q000379:methods*
D013723:Teratogens