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In vitro assessment human neurotoxicity and seizure liability using human induced pluripotent stem cells

 
 
Principle investigator: Anke Tukker, M.Sc 
Project leader: R.H.S. Westerink, Ph.D.
 
Project description:

The aim of this project is to develop, characterize and validate human induced pluripotent stem cell (hIPSC)-derived neurons grown in 2D and 3D for improved in vitro neurotoxicity testing.

 

Methods:

In this project, different hIPSC-derived neurons grown both in 2D and in 3D will be used to investigate the (neurotoxic) effects of selected reference compounds on neuronal function for comparison with existing in vitro (rodent) and in vivo (rodent and human) data. Therefore, effects of selected reference compounds on neuronal (electrical) activity and calcium homeostasis will be investigated with state of the art multi-electrode array recordings and single cell fluoresce microscopy, respectively. The overarching aim is the large-scale implementation of animal-free testing methods upon successful characterization and validation of truly predictive (2D or 3D) hIPSC-derived neurons for human neurotoxicity assessment.

Background:

Exposure to (environmental) chemicals may result in severe neurological effects and it is therefore of utmost importance to identify and understand the health risks associated with these exposures. Current test methods strongly rely on large-scale animal use, mainly rodents. Importantly, since rodents are not little humans, these tests are not truly predictive. Consequently, there is an urgent need for more predictive, human-based test methods, and to limit or even eliminate the use of animals for human neurotoxic hazard and risk assessment.

Within this project, specific expertise on cellular neurophysiological mechanisms and methodologies will therefore be combined with current state of the art (human) in vitro models grown in 2D and 3D to elucidate the suitability of the respective readouts and models for human neurotoxicity testing and risk assessment in vitro.

 

Recent publications:

- Tukker AM, Bouwman LMS, van Kleef RGDM, Hendriks HS, Legler J, Westerink RHS. Perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) acutely affect human α1β2γ2L GABAA receptor and spontaneous neuronal network function in vitro. Sci Rep. 10, 5311 (2020).

- Tukker AM, Van Kleef RGDM, Wijnolts FMJ, De Groot A, Westerink RHS. Towards animal-free neurotoxicity screening: Applicability of hiPSC-derived neuronal models for in vitro seizure liability assessment. ALTEX 37, 121-135 (2020).

- Tukker AM, Van Kleef RGDM, Wijnolts FMJ, De Groot A, Westerink RHS. Towards animal-free neurotoxicity screening: Applicability of hiPSC-derived neuronal models for in vitro seizure liability assessment. ALTEX [ahead of print]

- Tukker AM, Wijnolts FMJ, de Groot A, Wubbolts RW, Westerink RHS (2019). In vitro techniques for assessing neurotoxicity using human iPSC-derived neuronal models. NeuroMethods 145, 17-35.

- Tukker AM, Wijnolts FMJ, de Groot A, Westerink RHS. Human iPSC-derived neuronal models for in vitro neurotoxicity assessment. NeuroToxicology 67, 215-225 (2018).

- Tukker AM, de Groot MWGDM, Wijnolts FMJ, Kasteel EE, Hondebrink L, Westerink RHS (2016). Is the time right for in vitro neurotoxicity testing using human iPSC-derived neurons? ALTEX 33, 261-271.