Upscaling biological complexity to boost neuronal and oligodendroglia maturation and improve in vitro developmental neurotoxicity (DNT) evaluation

Prediction of in vivo prenatal chlorpyrifos exposure leading to developmental neurotoxicity in humans based on in vitro toxicity data by quantitative in vitro–in vivo extrapolation

“…The use of in silico methods such as PBK modelling serves as a predictive tool for any substance systemic exposure and could be valuable to support risk assessment when data are sparse, such as in pregnant women ( Coppola et al, 2021 ). In this work, we performed QIVIVE to evaluate the quantitative predictive value of an vitro test system for DNT effects which was assumed to be promising in this respect ( Pistollato et al, 2020 ; Pistollato et al, 2021 ; Nunes et al, 2022 ), using non-acetyl cholinesterase-dependent markers in hiPSC-derived neuronal/glial cell model.…”

Assessment of neurotransmitter release in human iPSC-derived neuronal/glial cells: a missing in vitro assay for regulatory developmental neurotoxicity testing

“…P3143) (0.1% solution in sodium-tetra-borate buffer) and laminin (mouse protein, 10 µg/mL) coated 24-well microelectrode array (MEA) plates (24-well glass MEA plate (24W300/30 G-288) V.232, from Multi-Channel System) in 100 µL volume, and after 20 min, 400 µL medium/well was added (50,000 cells/well in 0.5 mL). Further details about model characterization are provided in our previous studies [2] , [47] .…”

“…In our recently published paper, we demonstrated that human iPSC-derived neuronal cultures are suitable to measure neurotransmitter release [45] . Therefore, in the present study, we applied this HPLC technology to measure the release of neurotransmitters in a previously well characterized hiPSC-derived NSC culture undergoing differentiation towards a mixed population of neurons and glia [2] , [46] , [47] . We assessed glutamate release in control cultures and upon exposure to DL-threo-β-Hydroxyaspartic acid (DL-TBOA, a non-transportable glutamate uptake inhibitor [48] ), 4-aminopyridine (4-AP, which induces action potential-like membrane depolarization by blocking voltage-gated K + -channels, causing in vitro release of glutamate, [49] , [50] ), bafilomycin A1 (Bafilo, a specific and potent inhibitor of vacuolar-type H + -ATPase required for the exocytotic vesicle loading with glutamate [51] , [52] , [53] ), or a combination of 4-AP and Bafilo.…”

Quality criteria for in vitro human pluripotent stem cell-derived models of tissue-based cells

“…For example, it is hard to develop a co-culture of neurons and oligodendrocytes with active myelination, but differentiation towards myelinating oligodendrocytes can easily be achieved using 3D neurospheres, compensating for the limitation of using 2D monolayer cultures [57]. These 3D neurospheres enable robust differentiation towards both neuronal and glial cell populations, including myelination and formation of more mature neuronal network activity when compared to 2D monolayer culture [19,57,63].…”

“…Clearly, the selection of a 2D versus a 3D model is likely to affect the results obtained. For example, 2D models are still more suitable to assess neuronal functionality by analysis of spontaneous electrical activity using traditional multielectrode array platform [63]. However, the fast development of new technologies, and increasing number of new bioengineered devices more suitable for electrophysiological recordings in 3D [71][72][73][74] might change that.…”