Development and characterization of a human embryonic stem cell-derived 3D neural tissue model for neurotoxicity testing

Sandström, Jenny; Eggermann, Emmanuel; Charvet, Igor; Roux, Adrien; Toni, Nicolas; Greggio, Chiara; Broyer, A.; Monnet‐Tschudi, Florianne; Stoppini, Luc

doi:10.1016/j.tiv.2016.10.001

Cited by 40 publications

(34 citation statements)

References 62 publications

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“…The gene expression profiles of mouse and human ES cells have been proven to be distinct in, for example, several differentiation markers (Liu, Yin, & Faiola, 2017;Tandon & Jyoti, 2012;Wobus & Loser, 2011), which may lead to completely different responses to the test chemicals in molecular approaches (Yamane et al, 2016). Several recent studies have thus investigated the potential application of human embryonic stem cells (hESCs), associated with a variety of molecular endpoints, to predict the developmental toxicity in vitro (Ehashi, Suzuki, Ando, Sumida, & Saito, 2014;Hong & Jeung, 2013;Jung et al, 2014;Sandstrom et al, 2017). However, in addition to ethical issues, using hESCs for drug screening not only requires complex culturing conditions but also is costly and difficult to implement in the HT manner (Fang, Zhi, Yu, Lynch, & Jia, 2018;Rezvanfar, Hodjat, & Abdollahi, 2016).…”

Section: Discussionmentioning

confidence: 99%

An engineered mouse embryonic stem cell model with survivin as a molecular marker and EGFP as the reporter for high throughput screening of embryotoxic chemicals in vitro

Zang

Xin

Zhang

et al. 2019

Biotech & Bioengineering

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Embryonic stem cell test (EST) is the only generally accepted in vitro method for assessing embryotoxicity without animal sacrifice. However, the implementation and application of EST for regulatory embryotoxicity screening are impeded by its technical complexity, long testing period, and limited endpoint data. In this study, a high throughput embryotoxicity screening based on mouse embryonic stem cells (mESCs) expressing enhanced green fluorescent protein (EGFP) driven by a human survivin promoter and a human cytomegalovirus promoter, respectively, was developed. These EGFP expressing mESCs were cultured in three-dimensional (3D) fibrous scaffolds in microbioreactors on a multiwell plate with EGFP fluorescence signals as cell responses to chemicals monitored noninvasively in a high throughput manner. Nine chemicals with known developmental toxicity were used to validate the survivin-based embryotoxicity assay, which showed that strongly embryotoxic compounds such as 5-fluorouracil, retinoic acid, and methotrexate downregulated survivin expression by more than 50% in 3 days, while weakly embryotoxic compounds such as boric acid, methoxyacetic acid, and tetracyclin showed modest downregulation effect and nonembryotoxic saccharin, penicillin G, and acrylamide had negligible downregulation effect on survivin expression, confirming that survivin can be used as a molecular endpoint for high throughput screening of embryotoxicants. The potential developmental toxicity of three Chinese herbal medicines were also evaluated using this assay, demonstrating its application in in vitro developmental toxicity test for drug safety assessment.

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Section: Discussionmentioning

confidence: 99%

An engineered mouse embryonic stem cell model with survivin as a molecular marker and EGFP as the reporter for high throughput screening of embryotoxic chemicals in vitro

Zang

Xin

Zhang

et al. 2019

Biotech & Bioengineering

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“…There are also studies on more complex neural cell populations, such as neural rosette [ 68 , 69 ] and artificial neural constructs [ 50 , 70 ]. For example, highly consistent responses toward RA between in vitro neural rosette and in vivo systems confirmed the potentiality of neural rosette generation for developmental neurotoxicity tests [ 69 ].…”

Section: Major Achievements and Future Prospects Of Human Stem Cell Tmentioning

confidence: 99%

Prospects and Frontiers of Stem Cell Toxicology

Liu

Yin

Faiola

2017

Stem Cells and Development

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The development of stem cell biology has revolutionized regenerative medicine and its clinical applications. Another aspect through which stem cells would benefit human health is their use in toxicology. In fact, owing to their ability to differentiate into all the lineages of the human body, including germ cells, stem cells, and, in particular, pluripotent stem cells, can be utilized for the assessment, in vitro, of embryonic, developmental, reproductive, organ, and functional toxicities, relevant to human physiology, without employing live animal tests and with the possibility of high throughput applications. Thus, stem cell toxicology would tremendously assist in the toxicological evaluation of the increasing number of synthetic chemicals that we are exposed to, of which toxicity information is limited. In this review, we introduce stem cell toxicology, as an emerging branch of in vitro toxicology, which offers quick and efficient alternatives to traditional toxicology assessments. We first discuss the development of stem cell toxicology, and we then emphasize its advantages and highlight the achievements of human pluripotent stem cell-based toxicity research.

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Section: Introductionmentioning

confidence: 99%

“…In this study, we present minibrains, a brain spheroid organoid model generated by non-directed differentiation of neural stem cells derived from human iPSCs (NSC hiPS ) (Figure 1 and 2) 1 . By 6 weeks, minibrains display neuronal activity and by 12 weeks minibrains display synchronized neural networks (Figure 1) 1 . The size, simplicity and cost efficiency of generating minibrains make them an ideal choice for mass production for the purpose of large-scale screening and modelling studies ( Figure 2, Supplementary Table 1).…”

Section: Introductionmentioning

confidence: 99%

Mass Generation, Neuron Labelling and 3D Imaging of Minibrains

Govindan

Batti

Osterop

et al. 2020

Preprint

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Minibrain is a spherical in vitro 3D brain organoid model, composed of a mixed population of neurons and glial cells, generated from human iPSC derived neural stem cells. Despite the advances in human brain organoid models, there is a lack of labelling and imaging methodologies to characterize these models. In this study, we present a step-by-step methodology to generate human minibrain nurseries and novel strategies to subsequently label projection neurons, perform immunohistochemistry and 3D imaging of the minibrains at large multiplexable scales. To visualize projection neurons, we adapt viral transduction and to visualize the organization of cell types we implement immunohistochemistry. To facilitate 3D imaging of minibrains, we present here pipelines and accessories for one step mounting and clearing suitable for confocal microscopy. The pipelines are specifically designed in such a way that the assays can be multiplexed with ease for large-scale screenings using minibrains. Using the pipeline, we present i. dendrite morphometric properties obtained from 3D neuron morphology reconstructions and ii. distribution and quantification of cell types in 3D across whole mount organoids.

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Development and characterization of a human embryonic stem cell-derived 3D neural tissue model for neurotoxicity testing

Cited by 40 publications

References 62 publications

An engineered mouse embryonic stem cell model with survivin as a molecular marker and EGFP as the reporter for high throughput screening of embryotoxic chemicals in vitro

An engineered mouse embryonic stem cell model with survivin as a molecular marker and EGFP as the reporter for high throughput screening of embryotoxic chemicals in vitro

Prospects and Frontiers of Stem Cell Toxicology

Mass Generation, Neuron Labelling and 3D Imaging of Minibrains

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