2012
DOI: 10.2131/jts.37.845
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Assessment of technical protocols for novel embryonic stem cell tests with molecular markers (Hand1- and Cmya1-ESTs): a preliminary cross-laboratory performance analysis

Abstract: -The Hand1-and Cmya1-ESTs are novel short-term tests for embryotoxic chemicals using genetically engineering mouse ES cells for luciferase reporter gene assays. These ESTs allow convenient determination of differentiation toxicity and cell viability in a short duration with high throughput 96-well microplates for prediction of embryotoxicity of chemicals. To assess the Hand1-EST technical protocol, we firstly compared reporter gene assay and cytotoxicity test data for a representative compound (hydroxyurea) fr… Show more

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Cited by 15 publications
(6 citation statements)
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References 17 publications
(17 reference statements)
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“…Additionally, different cell lines such as H1, SA002 and ReliCell hES1 were used for other investigations (Adler et al , , ; Mehta et al , ). However, we observed the tendency that a IC 50 value for 5‐fluorouracil or indomethacin is similar to recent research by other groups using with mES and hES cells in ESTs methods (Genschow et al , ; Mehta et al , ; Suzuki et al , ). These results suggest that hES cells and the conditions of our experiments were appropriate for assessing the IC 50 values of embryotoxic chemicals within a relatively short period.…”
Section: Discussionsupporting
confidence: 92%
See 1 more Smart Citation
“…Additionally, different cell lines such as H1, SA002 and ReliCell hES1 were used for other investigations (Adler et al , , ; Mehta et al , ). However, we observed the tendency that a IC 50 value for 5‐fluorouracil or indomethacin is similar to recent research by other groups using with mES and hES cells in ESTs methods (Genschow et al , ; Mehta et al , ; Suzuki et al , ). These results suggest that hES cells and the conditions of our experiments were appropriate for assessing the IC 50 values of embryotoxic chemicals within a relatively short period.…”
Section: Discussionsupporting
confidence: 92%
“…Diversity related genetic factors, DNA methylation and DNA repair processes between experimental animals and humans have been reported in the development stage (Krtolica et al , ). Although EST using mES cells has routinely been used to evaluate embryotoxic chemicals (Spielmann et al , ; Suzuki et al , ), it has limits in incorrect classification of embryotoxic chemicals owing to species‐specific differences between humans and animals (Ginis et al , ).…”
Section: Introductionmentioning
confidence: 99%
“…While undifferentiated mouse ES cells have been used as ESTs, we evaluated embryotoxic chemicals in the vitro system using undifferentiated human ES cells because use of mouse ES cells has reached its limits due to interspecies distinction (53). When we modified the protocol for measurement of IC 50 values after exposure of embryotoxic chemicals for up to seven days, the IC 50 values of embryotoxic chemicals were similar to those reported by other groups using ESTs with mouse ES (14) and human ES cells (18,54). This result suggests that our system was appropriate for assessment of the IC 50 values of embryotoxic chemicals within a relatively short period.…”
Section: Developmental Toxicants On Undifferentiated Human Es Cellssupporting
confidence: 63%
“…The conclusion of the study was high predictability and accuracy with reduced test duration and manpower compared to the original EST. Furthermore, in 2012, we published another study (Suzuki et al, 2012) in which we investigated Hand1-and Cmya1-ESTs to explore reproducibility by comparing a set of 6 well-known test chemicals, including hydroxyurea, at four different laboratories. We managed to obtain good correspondence among all four laboratories, proving the high potency of transferability, intra-laboratory variability, and inter-laboratory variability of the protocols.…”
Section: Introductionmentioning
confidence: 99%