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|Title:||Protocol for the Differentiation of Human Induced Pluripotent Stem Cells into Mixed Cultures of Neurons and Glia for Neurotoxicity Testing|
|Authors:||PISTOLLATO FRANCESCA; CANOVAS JORDA DAVID; ZAGOURA DIMITRA; PRICE ANNA|
|Citation:||JOURNAL OF VISUALIZED EXPERIMENTS : JOVE no. 124 p. 1-14|
|Type:||Articles in periodicals and books|
|Abstract:||Human pluripotent stem cells are able to differentiate into various cell types that can be applied in human based in vitro toxicity assays. One major advantage is that the reprogramming of somatic cells to produce human induced pluripotent stem cells (hiPSCs) avoids the ethical and legislative issues related to the use of human embryonic stem cells (hESCs). HiPSCs can be expanded and efficiently differentiated into different types of neuronal and glial cells, serving as test systems for toxicity testing and, in particular, for the assessment of different pathways involved in neurotoxicity. Here we describe a protocol for the differentiation of hiPSCs into mixed cultures of neuronal and glial cells. We defined which signalling pathways are regulated and/or activated upon neuronal differentiation. Providing this information is critical in order to be able to apply the cell model for the new toxicity testing paradigm, in which chemicals are assessed based on their ability to perturb biological pathways. As a proof of concept we used rotenone, an inhibitor of mitochondrial respiratory complex I, to assess the activation of Nrf2 signaling pathway, a key regulator of the Antioxidant-Response-Element-(ARE)-driven cellular defense mechanism against oxidative stress.|
|JRC Directorate:||Health, Consumers and Reference Materials|
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