Single‐cell study of neural stem cells derived from human iPSCs reveals distinct progenitor populations with neurogenic and gliogenic potential

Lam, Matti; Sanosaka, Tsukasa; Lundin, Anders; Imaizumi, Kent; Etal, Damla; Karlsson, Fredrik; Clausen, Maryam; Cairns, Jonathan; Hicks, Ryan; Kohyama, Jun; Kele, Malin; Falk, Anna

doi:10.1111/gtc.12731

Cited by 26 publications

(37 citation statements)

References 29 publications

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“…NES C9 and C7 were biased towards neuronal commitment, while NES AF22 generated a mixture of glial and neuronal populations. Our data corroborate with findings from other studies; specifically, NES AF22 has been shown to produce a mixture of glial and neuronal population upon growth factor withdrawal(Lam, Sanosaka, et al ., 2019) while NES C7 mainly generated neurons(Lam, Moslem, et al ., 2019). Adhering to that paradigm, NES AF22 should have the greatest potential for astrocyte differentiation (compared to NES C7 and NES C9), but strikingly NES AF22 underperformed compared to NES C9 and NES C7.…”

Section: Discussionsupporting

confidence: 93%

“…Albeit having a different regional identity, all hiAstrocytes stained positive for the canonical astrocytic markers, S100B, AQP4, ALDH1L1 and CD44. Interestingly, the generated cells are GFAP + which was not observed in our previous protocol to generate astrocytes from long-term proliferating NES(Lundin et al ., 2018, 2020; Lam, Sanosaka, et al ., 2019). NES (all lines) did not stain for any of these markers but were positive for VIMENTIN (Figure S2).…”

Section: Discussionmentioning

confidence: 99%

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Generation of star-shaped human induced astrocytes with a mature inflammatory phenotype for CNS disease modeling

Voulgaris

Nikolakolpoulou

Herland

2021

Preprint

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Generating astrocytes from induced pluripotent stem cells has been hampered by either prolonged differentiation -spanning over two months -or by shorter protocols that generate immature astrocytes, devoid of salient inflammation-associated astrocytic traits pivotal for CNS neuropathological modeling. We directed human neural stem cells derived from induced pluripotent stem cells to astrocytic commitment and maturity by orchestrating an astrocytic-tuned culturing environment. In under 28 days, the generated cells express canonical and mature astrocytic markers, denoted by the expression of AQP4 and, remarkably, the expression and functionality of glutamate transporter EAAT2. We also show that this protocol generates astrocytes that encompass traits critical in CNS disease modeling, such as glutathione synthesis and secretion, upregulation of ICAM-1 and a cytokine secretion profile which is on par with primary astrocytes. This protocol generates a multifaceted astrocytic model suitable for CNS in vitro disease modeling and personalized medicine through brain-on-chip technologies.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Generation of star-shaped human induced astrocytes with a mature inflammatory phenotype for CNS disease modeling

Voulgaris

Nikolakolpoulou

Herland

2021

Preprint

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“…These additional factors help increase the abundance of GFAP + cells but may simultaneously induce aspects of astrocyte reactivity ( Sofroniew, 2020 ). Once directed toward an astroglial cell fate, some protocols allow time to mature astrocyte progenitors ( Lam et al, 2019b ) while others direct astrocyte maturation with additional exogenous molecules ( Krencik and Zhang, 2011 ). The former likely produces more bona fide astrocyte-like cells but at the consequence of longer protocols (up to 180 + days).…”

Section: D Glial Stem Cell Modelsmentioning

confidence: 99%

Growing Glia: Cultivating Human Stem Cell Models of Gliogenesis in Health and Disease

Lanjewar

Sloan

2021

Front. Cell Dev. Biol.

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Glia are present in all organisms with a central nervous system but considerably differ in their diversity, functions, and numbers. Coordinated efforts across many model systems have contributed to our understanding of glial-glial and neuron-glial interactions during nervous system development and disease, but human glia exhibit prominent species-specific attributes. Limited access to primary samples at critical developmental timepoints constrains our ability to assess glial contributions in human tissues. This challenge has been addressed throughout the past decade via advancements in human stem cell differentiation protocols that now offer the ability to model human astrocytes, oligodendrocytes, and microglia. Here, we review the use of novel 2D cell culture protocols, 3D organoid models, and bioengineered systems derived from human stem cells to study human glial development and the role of glia in neurodevelopmental disorders.

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“…For example, certain polymorphisms in the dyslexia candidate genes DYX1C1, DCDC2, and KIAA0319 correlate with white matter density in the brain of children [59]. Recently, protocols to differentiate NES cells into the [23,60]. We further focused our analysis on a group of DCGs highly replicated in genetic studies and previously linked to neuronal development and cilia.…”

Section: Discussionmentioning

confidence: 99%

Dyslexia Candidate Gene and Ciliary Gene Expression Dynamics During Human Neuronal Differentiation

et al. 2020

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Developmental dyslexia (DD) is a neurodevelopmental condition with complex genetic mechanisms. A number of candidate genes have been identified, some of which are linked to neuronal development and migration and to ciliary functions. However, expression and regulation of these genes in human brain development and neuronal differentiation remain uncharted. Here, we used human long-term self-renewing neuroepithelial stem (lt-NES, here termed NES) cells derived from human induced pluripotent stem cells to study neuronal differentiation in vitro. We characterized gene expression changes during differentiation by using RNA sequencing and validated dynamics for selected genes by qRT-PCR. Interestingly, we found that genes related to cilia were significantly enriched among upregulated genes during differentiation, including genes linked to ciliopathies with neurodevelopmental phenotypes. We confirmed the presence of primary cilia throughout neuronal differentiation. Focusing on dyslexia candidate genes, 33 out of 50 DD candidate genes were detected in NES cells by RNA sequencing, and seven candidate genes were upregulated during differentiation to neurons, including DYX1C1 (DNAAF4), a highly replicated DD candidate gene. Our results suggest a role of ciliary genes in differentiating neuronal cells and show that NES cells provide a relevant human neuronal model to study ciliary and DD candidate genes. Keywords Cilia. Ciliopathies. Reading disorder. Human neuroepithelial stem cells. RNA sequencing Abbreviations BBS Bardet-Biedl syndrome CAGE Cap analysis of gene expression DAVID Database for Annotation, Visualization and Integrated Discovery DCDC2 Doublecortin domain containing 2 DCG Dyslexia candidate gene DCX Doublecortin DD Developmental dyslexia DNAAF Dynein axonemal assembly factor DYX1C1 Dyslexia susceptibility 1 candidate 1 EGF Epidermal growth factor FANTOM Functional annotation of the mammalian genome FDR False discovery rate FGF Fibroblast growth factor Andrea Bieder and Masahito Yoshihara contributed equally to this work.

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Single‐cell study of neural stem cells derived from human iPSCs reveals distinct progenitor populations with neurogenic and gliogenic potential

Cited by 26 publications

References 29 publications

Generation of star-shaped human induced astrocytes with a mature inflammatory phenotype for CNS disease modeling

Generation of star-shaped human induced astrocytes with a mature inflammatory phenotype for CNS disease modeling

Growing Glia: Cultivating Human Stem Cell Models of Gliogenesis in Health and Disease

Dyslexia Candidate Gene and Ciliary Gene Expression Dynamics During Human Neuronal Differentiation

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