Generation of Bioengineered Neuronal Organoids (BENOs) from human pluripotent stem cells

Zafeiriou, Maria-Patapia; Bao, Guobin; Blenkle, Alica; Schreiber, Marie-Kristin; Zimmermann, Wolfram‐Hubertus

doi:10.21203/rs.3.pex-998/v1

Cited by 2 publications

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“…In a recent study, we used this approach to describe the effect of 4‐AP, a potassium channel blocker, on early neurogenesis in human cerebral organoids (Parmentier et al., 2022). Since the first description of cerebral organoids in mouse and humans, several protocols have been published both from embryonic stem cells and iPSCs (Birey et al., 2017; Eiraku et al., 2008; Giandomenico et al., 2019; Kadoshima et al., 2013; Lancaster et al., 2013; Paşca et al., 2015; Velasco et al., 2019; Yoon et al., 2019; Zafeiriou, Guobin, et al., 2020). Protocols differ mainly in the use of small molecules and growth factors to guide PSCs toward differentiating into regionally specific cell types (guided protocols) or approaches where PSCs progressively differentiate into the neuroectodermal lineage that is the default lineage in the absence of guiding signals (unguided protocols; Pașca et al., 2022).…”

Section: Commentarymentioning

confidence: 99%

Evaluation of Neurotoxicity With Human Pluripotent Stem Cell–Derived Cerebral Organoids

2023

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The recent development of human cerebral organoids provides an invaluable in vitro model of human brain development to assess the toxicity of natural or man-made toxic substances. By recapitulating key aspects of early human neurodevelopment, investigators can evaluate with this three-dimensional (3D) model the effect of certain compounds on the formation of neuronal networks and their electrophysiological properties with more physiological relevance than neurons grown in monolayers and in cultures composed of a unique cell type. This promising potential has contributed to the development of a large number of diverse protocols to generate human cerebral organoids, making interlaboratory comparisons of results difficult. Based on a previously published protocol to generate human cortical organoids (herein called cerebral organoids), we detail several approaches to evaluate the effect of chemicals on neurogenesis, apoptosis, and neuronal function when exogenously applied to cultured specimens. Here, we take as an example 4-aminopyridine, a potassium channel blocker that modulates the activity of neurons and neurogenesis, and describe a simple and cost-effective way to test the impact of this agent on cerebral organoids derived from human induced pluripotent stem cells. We also provide tested protocols to evaluate neurogenesis in cerebral organoids with ethynyl deoxyuridine labeling and neuronal activity with live calcium imaging and microelectrode arrays. Together, these protocols should facilitate the implementation of cerebral organoid technologies in laboratories wishing to evaluate the effects of specific compounds or conditions on the development and function of human neurons with only basic cell culture equipment.

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Section: Commentarymentioning

confidence: 99%

Evaluation of Neurotoxicity With Human Pluripotent Stem Cell–Derived Cerebral Organoids

2023

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Developmental GABA polarity switch and neuronal plasticity in Bioengineered Neuronal Organoids

et al. 2020

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Brain organoids are promising tools for disease modeling and drug development. For proper neuronal network formation excitatory and inhibitory neurons as well as glia need to co-develop. Here, we report the directed self-organization of human induced pluripotent stem cells in a collagen hydrogel towards a highly interconnected neuronal network at a macroscale tissue format. Bioengineered Neuronal Organoids (BENOs) comprise interconnected excitatory and inhibitory neurons with supportive astrocytes and oligodendrocytes. Giant depolarizing potential (GDP)-like events observed in early BENO cultures mimic early network activity of the fetal brain. The observed GABA polarity switch and reduced GDPs in >40 day BENO indicate progressive neuronal network maturation. BENOs demonstrate expedited complex network burst development after two months and evidence for long-term potentiation. The similarity of structural and functional properties to the fetal brain may allow for the application of BENOs in studies of neuronal plasticity and modeling of disease.

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Generation of Bioengineered Neuronal Organoids (BENOs) from human pluripotent stem cells

Abstract: Here we present a new directed differentiation and maturation protocol to generate Bioengineered Neuronal Organoids (BENOs) comprised of complex networks of inhibitory and excitatory neurons as well as supporting glial cells (astrocytes and myelinating oligodendrocytes).

Cited by 2 publications

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Evaluation of Neurotoxicity With Human Pluripotent Stem Cell–Derived Cerebral Organoids

Evaluation of Neurotoxicity With Human Pluripotent Stem Cell–Derived Cerebral Organoids

Developmental GABA polarity switch and neuronal plasticity in Bioengineered Neuronal Organoids

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