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Updated: Apr 19, 2026
Generation of iPSC-derived Human Brain Organoids to Model Early Neurodevelopmental Disorders
Published on: April 14, 2017
Distinct gene expression responses of two anticonvulsant drugs in a novel human embryonic stem cell based neural
Sjors H W Schulpen1, Esther de Jong1, Liset J J de la Fonteyne2
1Laboratory for Health Protection Research, National Institute for Public Health and the Environment (RIVM), Antonie van Leeuwenhoeklaan 9, 3721 MA Bilthoven, The Netherlands; Institute for Risk Assessment Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, 3584 CM Utrecht, The Netherlands.
Abstract:
Hazard assessment of chemicals and pharmaceuticals is increasingly gaining from knowledge about molecular mechanisms of toxic action acquired in dedicated in vitro assays. We have developed an efficient human embryonic stem cell neural differentiation test (hESTn) that allows the study of the molecular interaction of compounds with the neural differentiation process. Within the 11-day differentiation protocol of the assay, embryonic stem cells lost their pluripotency, evidenced by the reduced expression of stem cell markers Pou5F1 and Nanog. Moreover, stem cells differentiated into neural cells, with morphologically visible neural structures together with increased expression of neural differentiation-related genes such as βIII-tubulin, Map2, Neurogin1, Mapt and Reelin. Valproic acid (VPA) and carbamazepine (CBZ) exposure during hESTn differentiation led to concentration-dependent reduced expression of βIII-tubulin, Neurogin1 and Reelin. In parallel VPA caused an increased gene expression of Map2 and Mapt which is possibly related to the neural protective effect of VPA. These findings illustrate the added value of gene expression analysis for detecting compound specific effects in hESTn. Our findings were in line with and could explain effects observed in animal studies. This study demonstrates the potential of this assay protocol for mechanistic analysis of specific compound-induced inhibition of human neural cell differentiation.
Insights
A new human embryonic stem cell neural differentiation test (hESTn) effectively monitors chemical impacts on neural development. This assay reveals compound-specific effects on neural differentiation, aiding in hazard assessment.
Area of Science:
- Toxicology
- Developmental Biology
- Stem Cell Research
Background:
- Chemical and pharmaceutical hazard assessment benefits from understanding molecular toxicology mechanisms.
- In vitro assays are crucial for studying these mechanisms.
Purpose of the Study:
- To develop and validate an efficient human embryonic stem cell neural differentiation test (hESTn) for assessing compound interactions with neural differentiation.
- To investigate the utility of gene expression analysis within this assay for detecting compound-specific effects.
Main Methods:
- An 11-day human embryonic stem cell neural differentiation protocol (hESTn) was established.
- Stem cell pluripotency markers (Pou5F1, Nanog) and neural differentiation markers (βIII-tubulin, Map2, Neurogin1, Mapt, Reelin) were analyzed.
- The effects of Valproic acid (VPA) and Carbamazepine (CBZ) on neural differentiation were assessed via gene expression analysis.
Main Results:
- The hESTn assay successfully induced neural differentiation, confirmed by morphological changes and marker expression.
- VPA and CBZ exposure resulted in concentration-dependent inhibition of key neural differentiation markers (βIII-tubulin, Neurogin1, Reelin).
- VPA exposure showed increased expression of Map2 and Mapt, suggesting potential neuroprotective effects.
Conclusions:
- The hESTn assay is a valuable tool for mechanistic analysis of compound-induced inhibition of human neural cell differentiation.
- Gene expression analysis within the hESTn assay provides insights into compound-specific effects and potential mechanisms of action.
- Findings align with animal study observations, supporting the assay's relevance for hazard assessment.
