High throughput 3D gel-based neural organotypic model for cellular assays using fluorescence biosensors

Abstract

Three-dimensional (3D) organotypic models that capture native-like physiological features of tissues are being pursued as clinically predictive assays for therapeutics development. A range of these models are being developed to mimic brain morphology, physiology, and pathology of neurological diseases. Biofabrication of 3D gel-based cellular systems is emerging as a versatile technology to produce spatially and cell-type tailored, physiologically complex and native-like tissue models. Here we produce 3D fibrin gel-based functional neural co-culture models with human-iPSC differentiated dopaminergic or glutamatergic neurons and astrocytes. We further introduce genetically encoded fluorescence biosensors and optogenetics activation for real time functional measurements of intracellular calcium and levels of dopamine and glutamate neurotransmitters, in a high-throughput compatible plate format. We use pharmacological perturbations to demonstrate that the drug responses of 3D gel-based neural models are like those expected from in-vivo data, and in some cases, in contrast to those observed in the equivalent 2D neural models.

Advancing Towards Physiologically Relevant Models of the Brain: Three-Dimensional Human Induced Pluripotent Stem Cell (hiPSC)-Based Cell Culture Systems in Neuroscience

“…Co-culture models of neurons and astrocytes were established and advanced during the last years. The culturing of hiPSC-derived neurons and astrocytes in 3D systems includes scaffold-based 23 and scaffold-free systems 24 . As an example, Park and colleagues 24 generated a 3D spheroid model with hiPSCderived neurons and astrocytes which is reproducible in regard to spheroid size and cell type composition which is of advantage for high-throughput drug efficacy screenings.…”