Robust Generation of Person-Specific, Synchronously Active Neuronal Networks Using Purely Isogenic Human iPSC-3D Neural Aggregate Cultures

Izsák, Júlia; Seth, Henrik; Andersson, Mats; Vizlin‐Hodzic, Dzeneta; Theiss, Stephan; Hanse, Eric; Ågren, Hans; Funa, Keiko; Illes, Sebastian

doi:10.3389/fnins.2019.00351

Cited by 38 publications

(85 citation statements)

References 54 publications

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“…In the present study, we analyzed the functional and cellular impact of hCSF on hiPSC-3D neural aggregates (3D NAs) comprised of cortical neurons, glial cells, and residing proliferative neural cells ( Edri et al., 2015 , Izsak et al., 2019 ). For this purpose, we applied patch-clamp technique, microelectrode array (MEA) recordings, and confocal imaging on 3D NAs exposed to hCSF obtained from healthy adult individuals.…”

Section: Introductionmentioning

confidence: 99%

Human Cerebrospinal Fluid Promotes Neuronal Circuit Maturation of Human Induced Pluripotent Stem Cell-Derived 3D Neural Aggregates

et al. 2020

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Summary Human induced pluripotent stem cell (hiPSC)-derived in vitro neural and organoid models resemble fetal, rather than adult brain properties, indicating that currently applied cultivation media and supplements are insufficient to achieve neural maturation beyond the fetal stage. In vivo , cerebrospinal fluid molecules are regulating the transition of the immature fetal human brain into a mature adult brain. By culturing hiPSC-3D neural aggregates in human cerebrospinal fluid (hCSF) obtained from healthy adult individuals, we demonstrate that hCSF rapidly triggers neurogenesis, gliogenesis, synapse formation, neurite outgrowth, suppresses proliferation of residing neural stem cells, and results in the formation of synchronously active neuronal circuits in vitro within 3 days. Thus, a physiologically relevant and adult brain-like milieu triggers maturation of hiPSC-3D neural aggregates into highly functional neuronal circuits in vitro . The approach presented here opens a new avenue to identify novel physiological factors for the improvement of hiPSC neural in vitro models.

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Section: Introductionmentioning

confidence: 99%

Human Cerebrospinal Fluid Promotes Neuronal Circuit Maturation of Human Induced Pluripotent Stem Cell-Derived 3D Neural Aggregates

et al. 2020

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“…In the present study, five BD-iPSC lines and four control-iPSC lines were used for the generation of NSCs and mature 3D neural aggregates comprising of neurons, astrocytes, and oligodendroglia cells 12 (Fig. 1).…”

Section: Resultsmentioning

confidence: 99%

“…A copy of the detailed method description from Vizlin-Hodzic et al 8 can be found in the supplementary information. In the present study, five BD-iPSC lines and four control-iPSC lines were used for the generation of NSCs and mature 3D neural aggregates comprising of neurons, astrocytes, and oligodendroglia cells 12 (Fig. 1 ).…”

Section: Resultsmentioning

confidence: 99%

“…Human BNDF, GNDF, NT-3, FGF8, TGF-β, (20 ng/ ml each, all Peprotech) and DAPT (Tocris) were used as neurotrophic factors. For details about the generation and characterization of 3D neural aggregates see elsewhere 12 .…”

Section: Sample Selectionmentioning

confidence: 99%

“…For immunocytochemistry, hiPSC-NSCs and neural aggregates were cultured on glass coverslips. The protocol for immunocytochemistry is described in our previous study 12 . Neural stem cell cultures were stained with mouse Nestin (Abcam, ab22035, 1:500) antibody, and 3D neural aggregate cultures with mouse betaTubIII (R&D, MAB1195, 1:2000) to visualize neurons, rabbit GFAP (DAKO, now Labome, Z0334, 1:1000) to visualize astrocytes and mouse O4 (Abcam, ab53041, 1:200) to visualize oligodendroglial cells.…”

Section: Immunocytochemistry and Confocal Imagingmentioning

confidence: 99%

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Identification of candidate genetic variants and altered protein expression in neural stem and mature neural cells support altered microtubule function to be an essential component in bipolar disorder

et al. 2020

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Identification of causative genetic variants leading to the development of bipolar disorder (BD) could result in genetic tests that would facilitate diagnosis. A better understanding of affected genes and pathways is also necessary for targeting of genes that may improve treatment strategies. To date several susceptibility genes have been reported from genome-wide association studies (GWAS), but little is known about specific variants that affect disease development. Here, we performed quantitative proteomics and whole-genome sequencing (WGS). Quantitative proteomics revealed NLRP2 as the most significantly up-regulated protein in neural stem cells and mature neural cells obtained from BD-patient cell samples. These results are in concordance with our previously published transcriptome analysis. Furthermore, the levels of FEZ2 and CADM2 proteins were also significantly differentially expressed in BD compared to control derived cells. The levels of FEZ2 were significantly downregulated in neural stem cells (NSC) while CADM2 was significantly up-regulated in mature neuronal cell culture. Promising novel candidate mutations were identified in the ANK3, NEK3, NEK7, TUBB, ANKRD1, and BRD2 genes. A literature search of candidate variants and deregulated proteins revealed that there are several connections to microtubule function for the molecules putatively involved. Microtubule function in neurons is critical for axon structure and axonal transport. A functional dynamic microtubule is also needed for an advocate response to cellular and environmental stress. If microtubule dynamics is compromised by mutations, it could be followed by deregulated expression forming a possible explanation for the inherited vulnerability to stressful life events that have been proposed to trigger mood episodes in BD patients.

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Alginate‐Laminin Hydrogel Supports Long‐Term Neuronal Activity in 3D Human Induced Pluripotent Stem Cell‐Derived Neuronal Networks

Hartmann

Lauria

Bendt

et al. 2022

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For 3D neural cultures durable hydrogels are required, which persist over a long differentiation period and thus enable the maturation of neuronal networks (NN). Here, 3D models based on human induced pluripotent stem cell‐derived neural progenitor cells that are embedded in hydrogels of either pure alginate or alginate functionalized with the extracellular matrix protein laminin 111 (L111) are established. This study analyzes material characteristics such as porosity, L111 distribution and shear viscosity, cell compatibility of hydrogels by measuring viability and cytotoxicity, and neural function by monitoring cell migration, differentiation as well as NN formation and activity on multielectrode arrays. The addition of L111 increases neural migration and enhances differentiation into neurons and astrocytes as well as synaptogenesis in alginate hydrogels. NN formed in hydrogels are electrically active for up to 206 d and L111‐supplementation further increases electrical activity, network maturation, and synchronicity compared to 2D controls and NN grown in pure alginate hydrogels. L111 addition to alginate gels further accelerates recovery of electrical activity after blockage of sodium channels with tetrodotoxin. In conclusion, NN grown in alginate‐L111 hydrogel blends are promising models for future long‐term applications in disease modeling, drug or chemical evaluation.

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Robust Generation of Person-Specific, Synchronously Active Neuronal Networks Using Purely Isogenic Human iPSC-3D Neural Aggregate Cultures

Cited by 38 publications

References 54 publications

Human Cerebrospinal Fluid Promotes Neuronal Circuit Maturation of Human Induced Pluripotent Stem Cell-Derived 3D Neural Aggregates

Human Cerebrospinal Fluid Promotes Neuronal Circuit Maturation of Human Induced Pluripotent Stem Cell-Derived 3D Neural Aggregates

Identification of candidate genetic variants and altered protein expression in neural stem and mature neural cells support altered microtubule function to be an essential component in bipolar disorder

Alginate‐Laminin Hydrogel Supports Long‐Term Neuronal Activity in 3D Human Induced Pluripotent Stem Cell‐Derived Neuronal Networks

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