μNeurocircuitry: Establishing in vitro models of neurocircuits with human neurons

Abstract

Neurocircuits in the human brain govern complex behavior and involve connections from many different neuronal subtypes from different brain regions. Recent advances in stem cell biology have enabled the derivation of patient-specific human neuronal cells of various subtypes for the study of neuronal function and disease pathology. Nevertheless, one persistent challenge using these human-derived neurons is the ability to reconstruct models of human brain circuitry. To overcome this obstacle, we have developed a compartmentalized microfluidic device, which allows for spatial separation of cell bodies of different human-derived neuronal subtypes (excitatory, inhibitory and dopaminergic) but is permissive to the spreading of projecting processes. Induced neurons (iNs) cultured in the device expressed pan-neuronal markers and subtype specific markers. Morphologically, we demonstrate defined synaptic contacts between selected neuronal subtypes by synapsin staining. Functionally, we show that excitatory neuronal stimulation evoked excitatory postsynaptic current responses in the neurons cultured in a separate chamber.

Common Disease Is More Complex Than Implied by the Core Gene Omnigenic Model

“…We need high-throughput scale platforms to characterize vulnerable human genomes (i.e., ascertained as cases) for a battery of measures. Technological advances in cellular reprogramming now approach the costachievable possibility of integrating multiple disease-specific cell types in complex synthetic human tissues (Fantuzzo et al, 2017;Junaid et al, 2017;Wevers et al, 2016). These models can mimic disease pathology by integration with physiological or pathophysiological stressors, inflammatory cytokines and inflammatory cells, bacterial or viral challenges, or a wide range of experimental perturbations (chemical screens, CRISPR mutations, etc.).…”

Using human stem cells as a model system to understand the neural mechanisms of alcohol use disorders: Current status and outlook

“…Interestingly, a recent study reported that iN cells but not neurons derived from iPS cells maintained age related epigenetic marks (Mertens, Paquola, et al, 2015). The iN cells are generated by ectopic expression of proneuronal differentiation specific TFs (Pang et al, 2011;Vierbuchen et al, 2010) that allows generation of subtype specific human neuronal cells including excitatory (Zhang et al, 2013), inhibitory and dopaminergic (Fantuzzo et al, 2017) flavors. Thus, iN cell models may provide a viable model system that potentially maintains the epigenetic modifications contributing to the disease.…”

“…To address the complexity of human neural circuitry modeling, a multicompartmental microfluidic culturing systems have been developed, which allows different subtypes of human neurons to be cultured in different compartments, while permitting synaptic connectivity (Fantuzzo et al, 2017). Moreover, this system allows imaging as well as functional analysis.…”

Compartmentalized Devices as Tools for Investigation of Human Brain Network Dynamics

“…Electrophysiology and network activity measured by MEA revealed functional differences between the two cell populations. Compartmentalized devices open to patch clamp apparatus (Fantuzzo et al, ) or constructed on an MEA (Berdichevsky et al, ; Habibey et al, ) may provide an option for studies interested in electrophysiology within compartmentalized contexts.…”