Co-culture of Glutamatergic Neurons and Pediatric High-Grade Glioma Cells Into Microfluidic Devices to Assess Electrical Interactions.
Fiche publication
Date publication
novembre 2021
Journal
Journal of visualized experiments : JoVE
Auteurs
Membres identifiés du Cancéropôle Est :
Dr DONTENWILL Monique, Pr ENTZ-WERLE Natacha, Dr LHERMITTE Benoît
Tous les auteurs :
Fuchs Q, Batut A, Gleyzes M, Rontard J, Miny L, Libralato M, Vieira J, Debis D, Larramendy F, Honegger T, Messe M, Pierrevelcin M, Lhermitte B, Dontenwill M, Entz-Werlé N
Lien Pubmed
Résumé
Pediatric high-grade gliomas (pHGG) represent childhood and adolescent brain cancers that carry a rapid dismal prognosis. Since there is a need to overcome the resistance to current treatments and find a new way of cure, modeling the disease as close as possible in an in vitro setting to test new drugs and therapeutic procedures is highly demanding. Studying their fundamental pathobiological processes, including glutamatergic neuron hyperexcitability, will be a real advance in understanding interactions between the environmental brain and pHGG cells. Therefore, to recreate neurons/pHGG cell interactions, this work shows the development of a functional in vitro model co-culturing human-induced Pluripotent Stem (hiPS)-derived cortical glutamatergic neurons pHGG cells into compartmentalized microfluidic devices and a process to record their electrophysiological modifications. The first step was to differentiate and characterize human glutamatergic neurons. Secondly, the cells were cultured in microfluidic devices with pHGG derived cell lines. Brain microenvironment and neuronal activity were then included in this model to analyze the electrical impact of pHGG cells on these micro-environmental neurons. Electrophysiological recordings are coupled using multielectrode arrays (MEA) to these microfluidic devices to mimic physiological conditions and to record the electrical activity of the entire neural network. A significant increase in neuron excitability was underlined in the presence of tumor cells.
Référence
J Vis Exp. 2021 Nov 17;(177):