A multimodal 3D neuro-microphysiological system with neurite-trapping microelectrodes

Abstract Three-dimensional cell technologies as pre-clinical models are emerging tools for mimicking the structural and functional complexity of the nervous system. The accurate exploration of phenotypes in engineered 3D neuronal cultures, however, demands morphological, molecular and especially functional measurements. Particularly crucial is measurement of electrical activity of individual neurons with millisecond resolution. Current techniques rely on customized electrophysiological recording set-ups, characterized by limited throughput and poor integration with other readout modalities. Here we describe a novel approach, using multiwell glass microfluidic microelectrode arrays, allowing non-invasive electrical recording from engineered 3D neural tissues. We demonstrate parallelized studies with reference compounds, calcium imaging and optogenetic stimulation. Additionally, we show how microplate compatibility allows automated handling and high-content analysis of human induced pluripotent stem cell–derived neurons. This microphysiological platform opens up new avenues for high-throughput studies on the functional, morphological and molecular details of neurological diseases and their potential treatment by therapeutic compounds.

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A multimodal 3D neuro-microphysiological system with neurite-trapping microelectrodes

10.1101/2021.04.16.436793 ◽

2021 ◽

Author(s):

Beatriz Molina-Martinez ◽

Laura V Jentsch ◽

Fulya Ersoy ◽

Matthijs van der Moolen ◽

Stella Donato ◽

...

Keyword(s):

Neurological Diseases ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Electrophysiological Recording ◽

Neuronal Cultures ◽

Novel Approach ◽

Clinical Models ◽

Induced Pluripotent ◽

Therapeutic Compounds ◽

Automated Handling

Three-dimensional cell technologies as pre-clinical models are emerging tools mimicking the structural and functional complexity of the nervous system. The accurate exploration of phenotypes in engineered 3D neuronal cultures, however, demands morphological, molecular and especially functional measurements. Particularly crucial is measurement of electrical activity of individual neurons with millisecond resolution. Current techniques rely on customized electrophysiological recording set-ups, characterized by limited throughput and poor integration with other readout modalities. Here we describe a novel approach, using multiwell glass microfluidic microelectrode arrays, allowing non-invasive electrical recording from engineered 3D neural tissues. We demonstrate parallelized studies with reference compounds, calcium imaging and optogenetic stimulation. Additionally, we show how microplate compatibility allows automated handling and high-content analysis of human-induced pluripotent stem cell-derived neurons. This microphysiological platform opens up new avenues for high-throughput studies on the functional, morphological and molecular details of neurological diseases and their potential treatment by therapeutic compounds.

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Progress of Induced Pluripotent Stem Cell Technologies to Understand Genetic Epilepsy

International Journal of Molecular Sciences ◽

10.3390/ijms21020482 ◽

2020 ◽

Vol 21 (2) ◽

pp. 482 ◽

Cited By ~ 1

Author(s):

Bruno Sterlini ◽

Floriana Fruscione ◽

Simona Baldassari ◽

Fabio Benfenati ◽

Federico Zara ◽

...

Keyword(s):

Stem Cell ◽

Pluripotent Stem Cell ◽

Neurological Diseases ◽

Functional Characterization ◽

3D Structure ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Genetic Epilepsy ◽

Human Neurons ◽

Induced Pluripotent

The study of the pathomechanisms by which gene mutations lead to neurological diseases has benefit from several cellular and animal models. Recently, induced Pluripotent Stem Cell (iPSC) technologies have made possible the access to human neurons to study nervous system disease-related mechanisms, and are at the forefront of the research into neurological diseases. In this review, we will focalize upon genetic epilepsy, and summarize the most recent studies in which iPSC-based technologies were used to gain insight on the molecular bases of epilepsies. Moreover, we discuss the latest advancements in epilepsy cell modeling. At the two dimensional (2D) level, single-cell models of iPSC-derived neurons lead to a mature neuronal phenotype, and now allow a reliable investigation of synaptic transmission and plasticity. In addition, functional characterization of cerebral organoids enlightens neuronal network dynamics in a three-dimensional (3D) structure. Finally, we discuss the use of iPSCs as the cutting-edge technology for cell therapy in epilepsy.

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Hereditary Optic Neuropathies: Induced Pluripotent Stem Cell-Based 2D/3D Approaches

Genes ◽

10.3390/genes12010112 ◽

2021 ◽

Vol 12 (1) ◽

pp. 112

Author(s):

Marta García-López ◽

Joaquín Arenas ◽

M. Esther Gallardo

Keyword(s):

Stem Cell ◽

Retinal Ganglion Cells ◽

Pluripotent Stem Cell ◽

Ganglion Cells ◽

Genetic Disorders ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Underlying Mechanism ◽

Retinal Ganglion ◽

Induced Pluripotent

Inherited optic neuropathies share visual impairment due to the degeneration of retinal ganglion cells (RGCs) as the hallmark of the disease. This group of genetic disorders are caused by mutations in nuclear genes or in the mitochondrial DNA (mtDNA). An impaired mitochondrial function is the underlying mechanism of these diseases. Currently, optic neuropathies lack an effective treatment, and the implementation of induced pluripotent stem cell (iPSC) technology would entail a huge step forward. The generation of iPSC-derived RGCs would allow faithfully modeling these disorders, and these RGCs would represent an appealing platform for drug screening as well, paving the way for a proper therapy. Here, we review the ongoing two-dimensional (2D) and three-dimensional (3D) approaches based on iPSCs and their applications, taking into account the more innovative technologies, which include tissue engineering or microfluidics.

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Robust Expression of Functional NMDA Receptors in Human Induced Pluripotent Stem Cell-Derived Neuronal Cultures Using an Accelerated Protocol

Frontiers in Molecular Neuroscience ◽

10.3389/fnmol.2021.777049 ◽

2021 ◽

Vol 14 ◽

Author(s):

Jacob B. Ruden ◽

Mrinalini Dixit ◽

José C. Zepeda ◽

Brad A. Grueter ◽

Laura L. Dugan

Keyword(s):

Nmda Receptors ◽

Neural Progenitor Cells ◽

Induced Pluripotent Stem Cell ◽

Calcium Flux ◽

Neuronal Cultures ◽

Mature Neurons ◽

Health And Disease ◽

Nervous System Function ◽

Induced Pluripotent ◽

Human Ipsc

N-methyl-D-aspartate (NMDA) receptors are critical for higher-order nervous system function, but in previously published protocols to convert human induced pluripotent stem cells (iPSCs) to mature neurons, functional NMDA receptors (NMDARs) are often either not reported or take an extended time to develop. Here, we describe a protocol to convert human iPSC-derived neural progenitor cells (NPCs) to mature neurons in only 37 days. We demonstrate that the mature neurons express functional NMDARs exhibiting ligand-activated calcium flux, and we document the presence of NMDAR-mediated electrically evoked postsynaptic current. In addition to being more rapid than previous procedures, our protocol is straightforward, does not produce organoids which are difficult to image, and does not involve co-culture with rodent astrocytes. This could enhance our ability to study primate/human-specific aspects of NMDAR function and signaling in health and disease.

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Three-dimensional induced pluripotent stem-cell models of human brain angiogenesis

Microvascular Research ◽

10.1016/j.mvr.2020.104042 ◽

2020 ◽

Vol 132 ◽

pp. 104042 ◽

Cited By ~ 1

Author(s):

Raleigh M. Linville ◽

Diego Arevalo ◽

Joanna C. Maressa ◽

Nan Zhao ◽

Peter C. Searson

Keyword(s):

Stem Cell ◽

Human Brain ◽

Pluripotent Stem Cell ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Cell Models ◽

Induced Pluripotent ◽

Stem Cell Models ◽

Brain Angiogenesis

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From “Directed Differentiation” to “Neuronal Induction”: Modeling Neuropsychiatric Disease

Biomarker Insights ◽

10.4137/bmi.s20066 ◽

2015 ◽

Vol 10s1 ◽

pp. BMI.S20066 ◽

Cited By ~ 4

Author(s):

Seok-Man Ho ◽

Aaron Topol ◽

Kristen J. Brennand

Keyword(s):

Psychiatric Disorders ◽

Neurological Diseases ◽

Disease Onset ◽

Induced Pluripotent Stem Cell ◽

Directed Differentiation ◽

Neuropsychiatric Disease ◽

Somatic Cell Reprogramming ◽

Human Neurons ◽

Induced Pluripotent

Aberrant behavior and function of neurons are believed to be the primary causes of most neurological diseases and psychiatric disorders. Human postmortem samples have limited availability and, while they provide clues to the state of the brain after a prolonged illness, they offer limited insight into the factors contributing to disease onset. Conversely, animal models cannot recapitulate the polygenic origins of neuropsychiatric disease. Novel methods, such as somatic cell reprogramming, deliver nearly limitless numbers of pathogenic human neurons for the study of the mechanism of neuropsychiatric disease initiation and progression. First, this article reviews the advent of human induced pluripotent stem cell (hiPSC) technology and introduces two major methods, “directed differentiation” and “neuronal induction,” by which it is now possible to generate neurons for modeling neuropsychiatric disease. Second, it discusses the recent applications, and the limitations, of these technologies to in vitro studies of psychiatric disorders.

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