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Updated: Jun 17, 2026
Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation
Published on: June 17, 2015
Transcriptomics-based identification of developmental toxicants through their interference with cardiomyocyte
Dorien A M van Dartel1, Jeroen L A Pennings, Frederik J van Schooten
1Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands. dorien.van.dartel@rivm.nl
Abstract:
The embryonic stem cell test (EST) predicts developmental toxicity based on the inhibition of cardiomyocyte differentiation of embryonic stem cells (ESC). The subjective endpoint, the long culture duration together with the undefined applicability domain and related predictivity need further improvement to facilitate implementation of the EST into regulatory strategies. These aspects may be improved by studying gene expression changes in the ESC differentiation cultures and their modulation by compound exposure using transcriptomics. Here, we tested the developmental toxicants monobutyl phthalate and 6-aminonicotinamide. ESC were allowed to differentiated, and cardiomyocyte differentiation was assessed after 10 days of culture. RNA of solvent controls was collected after 0, 24, 48, 72 and 96 h of exposure, and RNA of developmental-toxicant-exposed cultures was collected after 24 and 96 h. Samples were hybridized to DNA microarrays, and 1355 genes were found differentially expressed among the unexposed experimental groups. These regulated genes were involved in differentiation-related processes, and Principal Component Analysis (PCA) based on these genes showed that the unexposed experimental groups appeared in chronological order in the PCA, which can therefore be regarded as a continuous representation of the differentiation track. The developmental-toxicant-exposed cultures appeared to deviate significantly from this differentiation track, which confirms the compound-modulating effects on the differentiation process. The incorporation of transcriptomics in the EST is expected to provide a more informative and improved endpoint in the EST as compared with morphology, allowing early detection of differentiation modulation. Furthermore, this approach may improve the definition of the applicability domain and predictivity of the EST.
Insights
This study enhances the embryonic stem cell test (EST) for predicting developmental toxicity by using transcriptomics to analyze gene expression changes during stem cell differentiation. This approach offers a more informative endpoint for improved toxicity testing.
Area of Science:
- Developmental toxicology
- Stem cell biology
- Transcriptomics
Background:
- The embryonic stem cell test (EST) predicts developmental toxicity by assessing cardiomyocyte differentiation inhibition.
- Current EST limitations include subjective endpoints, long culture times, and undefined applicability domains, hindering regulatory implementation.
- Transcriptomics offers a potential solution by analyzing gene expression changes during differentiation and compound exposure.
Purpose of the Study:
- To investigate the utility of transcriptomics for improving the EST.
- To analyze gene expression profiles during embryonic stem cell (ESC) differentiation and the effects of developmental toxicants.
- To assess if transcriptomics can provide a more objective and informative endpoint for the EST.
Main Methods:
- Embryonic stem cells (ESCs) were differentiated into cardiomyocytes over 10 days.
- Developmental toxicants (monobutyl phthalate, 6-aminonicotinamide) were applied.
- RNA was collected at various time points and analyzed using DNA microarrays.
- Principal Component Analysis (PCA) was used to visualize gene expression patterns.
Main Results:
- 1355 differentially expressed genes were identified in unexposed ESCs, reflecting differentiation processes.
- PCA demonstrated a chronological differentiation track for unexposed cells.
- Toxicant-exposed cultures significantly deviated from the normal differentiation track, indicating compound-induced modulation.
- Transcriptomics revealed compound-specific effects on differentiation pathways.
Conclusions:
- Incorporating transcriptomics into the EST provides a more informative endpoint than traditional morphology.
- This approach allows for early detection of differentiation modulation by developmental toxicants.
- Transcriptomics may enhance the applicability domain and predictivity of the EST for regulatory use.
