Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation

Zhou, Shuling; Szczesna, Karolina; Ochałek, Anna; Kobolák, Julianna; Varga, Eszter; Nemes, Csilla; Chandrasekaran, Abinaya; Rasmussen, Mette; Cirera, Susanna; Hyttel, Poul; Dinnyés, András; Freude, Kristine; Avci, Hasan X.

doi:10.1155/2016/4937689

Cited by 59 publications

(50 citation statements)

References 61 publications

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“…Interestingly, serious attempts have been made to study the employment and the differences between 2D or 3D cultures 28,29 . Zhou et al demonstrated that culturing induced pluripotent stem cells (iPSCs) in 3D conditions resulted in higher expression of both neural progenitor (Pax6 and NESTIN) and astrocyte (GFAP and AQP4) markers as well as yielding a more homogenous cell population compared to those iPSCs cultured in 2D fashion 30 . Recently, András Dinnyés's team reported that iPSCs neural induction in 3D produced a higher number of neural progenitor cells and neurons with longer neurites 29 .…”

Section: Discussionmentioning

confidence: 99%

Generating homogenous cortical preplate and deep-layer neurons using a combination of 2D and 3D differentiation cultures

Alsanie

Bahri

Habeeballah

et al. 2020

Sci Rep

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Embryonic stem cells (ESCs) can be used to derive different neural subtypes. Current differentiation protocols generate heterogeneous neural subtypes rather than a specific neuronal population. Here, we present a protocol to derive separate two-deep layer cortical neurons from mouse eScs (meScs). mESCs were differentiated into mature Tbr1 or Ctip2-positive neurons using a monolayer-based culture for neural induction and neurosphere-based culture for neural proliferation and expansion. the differentiation protocol relies on SMAD inhibition for neural induction and the use of FGF2 and EGF for proliferation and it is relatively short as mature neurons are generated between differentiation days 12-16. Compared with the monolayer-based differentiation method, mESCs can be directed to generate specific deep-layer cortical neurons rather than heterogeneous cortical neurons that are generated using the monolayer differentiation culture. The early analysis of progenitors using flow cytometry, immunocytochemistry, and qRT-PCR showed high neuralization efficiency. The immunocytochemistry and flow cytometry analyses on differentiation days 12 and 16 showed cultures enriched in Tbr1-and Ctip2-positive neurons, respectively. Conversely, the monolayer differentiation culture derived a mixture of Tbr1 and Ctip2 mature neurons. Our findings suggested that implementing a neurospherebased culture enabled directing neural progenitors to adopt a specific cortical identity. The generated progenitors and neurons can be used for neural-development investigation, drug testing, disease modelling, and examining novel cellular replacement therapy strategies. Embryonic stem cells (ESCs) are an invaluable research tool as they can differentiate into the three germ layers in vitro. The advancement in the differentiation protocols has enabled the derivation of different neural progenitors and mature neurons. These neurons can be used in disease modelling, drug testing, modelling development in a dish, and in establishing novel strategies for cellular replacement therapies. Although the interest has been shifted towards using human embryonic stem cells (hESCs), mouse embryonic stem cells (mESCs) still have a great potential in the field. Essentially, mESCs can be genetically manipulated where their differentiation is

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Section: Discussionmentioning

confidence: 99%

Generating homogenous cortical preplate and deep-layer neurons using a combination of 2D and 3D differentiation cultures

Alsanie

Bahri

Habeeballah

et al. 2020

Sci Rep

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“…The generation of cell types such as EBs, neural rosettes, and NPCs mimics various stages of in utero neurogenesis 21 . Few protocols also generated unconventional cell types such as EZ spheres 202 or neurospheres 205 . Such alternatives aim to eliminate laborious differentiation steps involving EB formation and manual picking of rosettes.…”

Section: Generating Relevant Neuronal Cell Types For Pdmentioning

confidence: 99%

Genetic predispositions of Parkinson’s disease revealed in patient-derived brain cells

Tran

Anastacio

Bardy

2020

npj Parkinsons Dis.

111

103

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Parkinson's disease (PD) is the second most prevalent neurological disorder and has been the focus of intense investigations to understand its etiology and progression, but it still lacks a cure. Modeling diseases of the central nervous system in vitro with human induced pluripotent stem cells (hiPSC) is still in its infancy but has the potential to expedite the discovery and validation of new treatments. Here, we discuss the interplay between genetic predispositions and midbrain neuronal impairments in people living with PD. We first summarize the prevalence of causal Parkinson's genes and risk factors reported in 74 epidemiological and genomic studies. We then present a meta-analysis of 385 hiPSC-derived neuronal lines from 67 recent independent original research articles, which point towards specific impairments in neurons from Parkinson's patients, within the context of genetic predispositions. Despite the heterogeneous nature of the disease, current iPSC models reveal converging molecular pathways underlying neurodegeneration in a range of familial and sporadic forms of Parkinson's disease. Altogether, consolidating our understanding of robust cellular phenotypes across genetic cohorts of Parkinson's patients may guide future personalized drug screens in preclinical research.

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“…Initially generated using rodent embryonic stem cells (Eiraku et al, ), recent advancements in human stem cell culture have enabled the generation of 3D organoids from pluripotent human embryonic stem cells, multipotent somatic stem cells, and induced‐pluripotent stem cells (iPSCs) allowing expansion directly from patient tissue (Lancaster et al, ). In addition, strategies for reprogramming iPS cells into all major cell types of the nervous system including neurons (Hunsberger et al, ), oligodendrocytes (Prasad et al, ), astrocytes (Jiang et al, ; Zhou et al, ), and microglia (Muffat et al, ) have been developed providing an invaluable resource for studying human neurological disease. Using these techniques, protocols for the generation of human brain‐like cerebral (Lancaster et al, ), cerebellar (Muguruma, Nishiyama, Kawakami, Hashimoto, & Sasai, ), forebrain (Qian et al, ), and mid‐brain (Jo et al, ; Monzel et al, ) organoids have been reported.…”

Section: Cutting Edge Technologies To Understand Glial Function Cellmentioning

confidence: 99%

Drug discovery for remyelination and treatment of MS

Cole

Early

Lyons

2017

Glia

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Glia constitute the majority of the cells in our nervous system, yet there are currently no drugs that target glia for the treatment of disease. Given ongoing discoveries of the many roles of glia in numerous diseases of the nervous system, this is likely to change in years to come. Here we focus on the possibility that targeting the oligodendrocyte lineage to promote regeneration of myelin (remyelination) represents a therapeutic strategy for the treatment of the demyelinating disease multiple sclerosis, MS. We discuss how hypothesis driven studies have identified multiple targets and pathways that can be manipulated to promote remyelination in vivo, and how this work has led to the first ever remyelination clinical trials. We also highlight how recent chemical discovery screens have identified a host of small molecule compounds that promote oligodendrocyte differentiation in vitro. Some of these compounds have also been shown to promote myelin regeneration in vivo, with one already being trialled in humans. Promoting oligodendrocyte differentiation and remyelination represents just one potential strategy for the treatment of MS. The pathology of MS is complex, and its complete amelioration may require targeting multiple biological processes in parallel. Therefore, we present an overview of new technologies and models for phenotypic analyses and screening that can be exploited to study complex cell-cell interactions in in vitro and in vivo systems. Such technological platforms will provide insight into fundamental mechanisms and increase capacities for drug-discovery of relevance to glia and currently intractable disorders of the CNS.

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Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation

Cited by 59 publications

References 61 publications

Generating homogenous cortical preplate and deep-layer neurons using a combination of 2D and 3D differentiation cultures

Generating homogenous cortical preplate and deep-layer neurons using a combination of 2D and 3D differentiation cultures

Genetic predispositions of Parkinson’s disease revealed in patient-derived brain cells

Drug discovery for remyelination and treatment of MS

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