Hand1-Luc embryonic stem cell test (Hand1-Luc EST): a novel rapid and highly reproducible in vitro test for embryotoxicity by measuring cytotoxicity and differentiation toxicity using engineered mouse ES cells

The Journal of Toxicological Sciences
Florian Le CozKoichi Saito

Abstract

The embryonic stem cell test (EST) is a promising alternative method for evaluating embryotoxicity of test chemicals by measuring cytotoxicity and differentiation toxicity using mouse ES cells. Differentiation toxicity is analyzed by microscopically counting the beating of embryonic bodies after 10 days of culture. However, improvements are necessary to reduce the laborious manipulations involved and the time required to obtain results. We have previously reported the successful stable transfection of ES cells (ES-D3) with the heart and neural crest derivatives expressed transcript 1 (Hand1) gene and the establishment of a 96-well multi-plate-based new EST with luciferase reporter assay 6 days after treatment with test chemicals. Now, we propose an even more rapid and easier EST, named Hand1-Luc EST. We established another cell line to monitor the Hand1 gene expression via a luciferase reporter gene. By mRNA analysis and luciferase assay, we examined in detail the luciferase activity during cell differentiation, which allowed us to reduce the time of measurement from day 6 to day 5 (120 hr). Furthermore, the protocol was improved, with, among others, the measurement of cytotoxicity and differentiation toxicity taking place in t...Continue Reading

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Citations

Sep 7, 2017·Stem Cells and Development·Shuyu LiuFrancesco Faiola
Sep 1, 2018·Toxicological Sciences : an Official Journal of the Society of Toxicology·Anthony L Luz, Erik J Tokar
Feb 20, 2020·Toxicological Sciences : an Official Journal of the Society of Toxicology·Todd J ZurlindenThomas B Knudsen
Feb 10, 2018·Luminescence : the Journal of Biological and Chemical Luminescence·Katsuhiro UnoYoshihiro Nakajima
Feb 9, 2018·Nihon yakurigaku zasshi. Folia pharmacologica Japonica·Koichi SaitoKumiko Kobayashi
Sep 15, 2017·Toxicological Sciences : an Official Journal of the Society of Toxicology·Kumiko KobayashiKoichi Saito

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