Human induced pluripotent stem cell (hiPSC)-derived neurons respond to convulsant drugs when co-cultured with hiPSC-derived astrocytes

Ishii, M; Yamamoto, Kōji; Shoji, Masanobu; Asami, Asano; Kawamata, Yuji

doi:10.1016/j.tox.2017.06.010

Cited by 55 publications

(55 citation statements)

References 34 publications

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“…Our data suggest that the majority of our cells are spinal cord motor neurons (positive for ChAT and, to a lesser extent, for ISL1/2), expressing appropriate neurotransmitter receptors, with AMPA receptors being more represented than GABA and glycine receptors, as would be expected in the spinal cord. Importantly, in accordance with our protocol of ventralization and caudalization, and in contrast to previous studies involving hiPSC-cortical neurons 8,9 , only a very small percentage of neurons were corticospinal CTIP2 + motor neurons.…”

Section: Discussionsupporting

confidence: 75%

“…The few previously published studies on MEA recording from co-cultures of hiPSC-A/hiPSC-N have been carried out using human iPSC-derived cortical neurons 5, 8,9 . Consistent with our observations, these studies have demonstrated that human astrocytes enhance different MEA parameters including: spiking rate, bursting rate and number of spiking and bursting electrodes.…”

Section: Discussionmentioning

confidence: 99%

“…Previous MEA studies have focused on hiPSC-derived cortical neurons, either alone [4][5][6] or in co-culture with primary rodent cortical astrocytes 7 and more recently with hiPSC-derived astrocytes 5, 8,9,10 . Despite an established body of evidence suggesting that astrocytes contribute to neuronal electrophysiological maturation by promoting synaptogenesis 11 , MEA-and human iPSCbased paradigms, still influenced by a traditionally neuron-centered perspective, have not yet been utilized to model this unique glial contribution.…”

Section: Introductionmentioning

confidence: 99%

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Role of human induced pluripotent stem cell-derived spinal cord astrocytes in the functional maturation of motor neurons in a multielectrode array system

Taga

Dastgheyb

Habela

et al. 2019

Preprint

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The ability to generate human induced pluripotent stem cell (hiPSC)-derived neural cells displaying region-specific phenotypes is of particular interest for modeling central nervous system (CNS) biology in vitro. We describe a unique method by which spinal cord hiPSC-derived astrocytes (hiPSC-A) are cultured with spinal cord hiPSC-derived motor neurons (hiPSC-MN) in a multielectrode array (MEA) system to record electrophysiological activity over time. We show that hiPSC-A enhance hiPSC-MN electrophysiological maturation in a time-dependent fashion.The sequence of plating, density, and age in which hiPSC-As are co-cultured with MN, but not their respective hiPSC line origin, are factors that influence neuronal electrophysiology. When compared to co-culture with mouse primary spinal cord astrocytes, we observe an earlier and more robust electrophysiological maturation in the fully human cultures, suggesting that the human origin is relevant to the recapitulation of astrocyte/motor neuron cross-talk. Finally, we test pharmacological compounds on our MEA platform and observe changes in electrophysiological activity which confirm hiPSC-MN maturation. These findings are supported by immunocytochemistry and real time PCR studies in parallel cultures demonstrating human astrocyte mediated changes in the structural maturation and protein expression profiles of the neurons. Interestingly, this relationship is reciprocal and co-culture with neurons influences astrocyte maturation as well. Taken together these data indicate that in a human in vitro spinal cord culture system, astrocytes alter hiPSC-MN maturation in a time-dependent and species specific manner and suggest a closer approximation of in vivo conditions. Main Points1. We developed a method for the co-culture of human iPSC-A/MN for multielectrode array recordings.2. The morphological, molecular, pharmacological, and electrophysiological characterization of the co-cultures suggests bidirectional maturation.

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Section: Discussionsupporting

confidence: 75%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Role of human induced pluripotent stem cell-derived spinal cord astrocytes in the functional maturation of motor neurons in a multielectrode array system

Taga

Dastgheyb

Habela

et al. 2019

Preprint

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“…The regional specificity of this paradigm is particularly relevant in precision medicine strategies for modeling neurological diseases. Previous literature on MEA applied to hiPSC studies have relied exclusively upon hiPSC‐derived cortical neurons, for in vitro modeling of epilepsy and seizures . Although recent MEA studies have used neurons cocultured with human iPSC‐derived astrocytes , less attention has been given to how astrocytes may influence neuronal electrophysiological activity—particularly in the context of human biology.…”

Section: Discussionmentioning

confidence: 99%

“…Previous MEA studies have focused on hiPSC‐derived cortical neurons, either alone or in coculture with primary rodent cortical astrocytes and more recently with hiPSC‐derived astrocytes . Despite an established body of evidence suggesting that astrocytes contribute to neuronal electrophysiological maturation by promoting synaptogenesis , MEA‐ and hiPSC‐based paradigms, still influenced by a traditionally neuron‐centered perspective, have not yet been used to model this unique glial contribution.…”

Section: Introductionmentioning

confidence: 99%

Role of Human-Induced Pluripotent Stem Cell-Derived Spinal Cord Astrocytes in the Functional Maturation of Motor Neurons in a Multielectrode Array System

Taga

Dastgheyb

Habela

et al. 2019

Stem Cells Translational Medicine

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The ability to generate human-induced pluripotent stem cell (hiPSC)-derived neural cells displaying region-specific phenotypes is of particular interest for modeling central nervous system biology in vitro. We describe a unique method by which spinal cord hiPSC-derived astrocytes (hiPSC-A) are cultured with spinal cord hiPSC-derived motor neurons (hiPSC-MN) in a multielectrode array (MEA) system to record electrophysiological activity over time. We show that hiPSC-A enhance hiPSC-MN electrophysiological maturation in a time-dependent fashion. The sequence of plating, density, and age in which hiPSC-A are cocultured with MN, but not their respective hiPSC line origin, are factors that influence neuronal electrophysiology. When compared to coculture with mouse primary spinal cord astrocytes, we observe an earlier and more robust electrophysiological maturation in the fully human cultures, suggesting that the human origin is relevant to the recapitulation of astrocyte/motor neuron crosstalk. Finally, we test pharmacological compounds on our MEA platform and observe changes in electrophysiological activity, which confirm hiPSC-MN maturation. These findings are supported by immunocytochemistry and real-time PCR studies in parallel cultures demonstrating human astrocyte mediated changes in the structural maturation and protein expression profiles of the neurons. Interestingly, this relationship is reciprocal and coculture with neurons influences astrocyte maturation as well. Taken together, these data indicate that in a human in vitro spinal cord culture system, astrocytes support hiPSC-MN maturation in a time-dependent and species-specific manner and suggest a closer approximation of in vivo conditions. STEM CELLS TRANSLATIONAL MEDICINE 2019;8:1272-1285 SIGNIFICANCE STATEMENTThis study develops a method by which human-induced pluripotent stem cell-derived astrocytes (hiPSC-A) with distinct spinal cord identity are cocultured with spinal cord motor neurons (hiPSC-MN) for multielectrode array (MEA) recordings. It also demonstrates that hiPSC-A influence the morphological, molecular, and electrophysiological maturation of hiPSC-MN. Similarly, this study shows that hiPSC-A maturation is enhanced by the coculture with hiPSC-MN. This fully human, spinal cord-specific, coculture platform with MEA analyses provides a new tool for investigating astrocyte/MN interactions and has the potential to more accurately model human diseases with spinal cord pathology, including spinal muscular atrophy and amyotrophic lateral sclerosis.

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Progress in the molecular mechanisms of genetic epilepsies using patient‐induced pluripotent stem cells

et al. 2018

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SummaryResearch findings on the molecular mechanisms of epilepsy almost always originate from animal experiments, and the development of induced pluripotent stem cell (iPSC) technology allows the use of human cells with genetic defects for studying the molecular mechanisms of genetic epilepsy (GE) for the first time. With iPSC technology, terminally differentiated cells collected from GE patients with specific genetic etiologies can be differentiated into many relevant cell subtypes that carry all of the GE patient's genetic information. iPSCs have opened up a new research field involving the pathogenesis of GE. Using this approach, studies have found that gene mutations induce GE by altering the balance between neuronal excitation and inhibition, which is associated. among other factors, with neuronal developmental disturbances, ion channel abnormalities, and synaptic dysfunction. Simultaneously, astrocyte activation, mitochondrial dysfunction, and abnormal signaling pathway activity are also important factors in the molecular mechanisms of GE.

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Human induced pluripotent stem cell (hiPSC)-derived neurons respond to convulsant drugs when co-cultured with hiPSC-derived astrocytes

Cited by 55 publications

References 34 publications

Role of human induced pluripotent stem cell-derived spinal cord astrocytes in the functional maturation of motor neurons in a multielectrode array system

Role of human induced pluripotent stem cell-derived spinal cord astrocytes in the functional maturation of motor neurons in a multielectrode array system

Role of Human-Induced Pluripotent Stem Cell-Derived Spinal Cord Astrocytes in the Functional Maturation of Motor Neurons in a Multielectrode Array System

Progress in the molecular mechanisms of genetic epilepsies using patient‐induced pluripotent stem cells

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