Differentiation of neural rosettes from human pluripotent stem cells in vitro is sequentially regulated on a molecular level and accomplished by the mechanism reminiscent of secondary neurulation

Fedorová, Veronika; Váňová, Tereza; Elrefae, Lina Mohamed Bahaael; Pospíšil, Jakub; Petrášová, Martina; Kolajova, Veronika; Hudáčová, Zuzana; Baniariova, Jana; Barak, Manouchehr; Pešková, Lucie; Bárta, Tomáš; Kaucká, Markéta; Killinger, Michael; Večeřa, Josef; Bernatík, Ondřej; Čajánek, Lukáš; Hříbková, Hana; Bohačiaková, Dáša

doi:10.1016/j.scr.2019.101563

Cited by 59 publications

(63 citation statements)

References 45 publications

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“…Neural induction of PSCs may be conducted in 2D conditions (monolayer) (Chambers et al, 2009;Patani et al, 2009;Fedorova et al, 2019) or in 3D conditions (embryoid bodies) (Figure 1) (Watanabe et al, 2007;Eiraku et al, 2008). At the same time, further generation and cultivation of NSCs can be achieved in several ways: 2D monoculture (Patani et al, 2009;Fedorova et al, 2019), 2D neural rosettes (Watanabe et al, 2007;Elkabetz et al, 2008;Chambers et al, 2009), through 3D cultures (Eiraku et al, 2008;Mariani et al, 2012), or in organoids (Lancaster et al, 2013;Pasxca et al, 2015;Qian et al, 2016).…”

Section: Protocolsmentioning

confidence: 99%

“…They also found that the addition of N2 and B27 supplements to serum-free culture media increased the efficiency of neural differentiation. Several experiments have demonstrated neural differentiation of ESCs and iPSCs using monolayer culture in the presence of N2 and B27, with (Ying et al, 2003;Lin et al, 2010;Matulka et al, 2013), and without FGF-2 (Lippmann et al, 2014;Fedorova et al, 2019).…”

Section: Default Differentiation Protocolsmentioning

confidence: 99%

“…Despite the variety of protocols for obtaining NSCs using various morphogens and inhibitors (Zhang et al, 2001;Perrier et al, 2004;Watanabe et al, 2007;Chambers et al, 2009;Koch et al, 2009;Li et al, 2011), it has turned out that they are not necessary for the successful initiation of neural differentiation of PSCs. Based on the 2D culture system, a protocol with a high efficiency (>95%) of NSC generation from hiPSCs in a serum-free culture medium, with low cell density plating, and without any morphogens or factors (FGF-2, BMP-inhibitors and others) was developed (Fedorova et al, 2019). This method enables obtaining a homogenous Pax6/Sox1 positive population of NSCs in 6 days of hiPCS cultivation in the differentiation medium.…”

Section: Dual-smad-inhibitionmentioning

confidence: 99%

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Neural Stem Cells and Methods for Their Generation From Induced Pluripotent Stem Cells in vitro

Galiakberova

Дашинимаев

2020

Front. Cell Dev. Biol.

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Neural stem cells (NSCs) provide promising approaches for investigating embryonic neurogenesis, modeling of the pathogenesis of diseases of the central nervous system, and for designing drug-screening systems. Such cells also have an application in regenerative medicine. The most convenient and acceptable source of NSCs is pluripotent stem cells (embryonic stem cells or induced pluripotent stem cells). However, there are many different protocols for the induction and differentiation of NSCs, and these result in a wide range of neural cell types. This review is intended to summarize the knowledge accumulated, to date, by workers in this field. It should be particularly useful for researchers who are beginning investigations in this area of cell biology.

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Section: Protocolsmentioning

confidence: 99%

Section: Default Differentiation Protocolsmentioning

confidence: 99%

Section: Dual-smad-inhibitionmentioning

confidence: 99%

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Neural Stem Cells and Methods for Their Generation From Induced Pluripotent Stem Cells in vitro

Galiakberova

Дашинимаев

2020

Front. Cell Dev. Biol.

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“…This results in emergence of a singular neuroepithelial tube that spans the entire rostrocaudal axis of the embryo's dorsal plane and serves as the foundation of all CNS tissues. Human pluripotent stem cells can readily differentiate into neuroepithelial cells that spontaneously polarize to form neural rosettes 55 , which are thought to recapitulate critical features of the early embryonic neural tube's structure and function 56,57 . Further, neural rosettes can give rise to extensive levels of human corticogenesis.…”

Section: Bax/bak Dko Hipsc-derived Neural Rosettes Show Larger Lumen mentioning

confidence: 99%

Modeling the function of BAX and BAK in early human brain development using iPSC-derived systems

et al. 2020

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Intrinsic apoptosis relies on the ability of the BCL-2 family to induce the formation of pores on the outer mitochondrial membrane. Previous studies have shown that both BAX and BAK are essential during murine embryogenesis, and reports in human cancer cell lines identified non-canonical roles for BAX and BAK in mitochondrial fission during apoptosis. BAX and BAK function in human brain development remains elusive due to the lack of appropriate model systems. Here, we generated BAX/BAK double knockout human-induced pluripotent stem cells (hiPSCs), hiPSC-derived neural progenitor cells (hNPCs), neural rosettes, and cerebral organoids to uncover the effects of BAX and BAK deletion in an in vitro model of early human brain development. We found that BAX and BAK-deficient cells have abnormal mitochondrial morphology and give rise to aberrant cortical structures. We suggest crucial functions for BAX and BAK during human development, including maintenance of homeostatic mitochondrial morphology, which is crucial for proper development of progenitors and neurons of the cortex. Human pluripotent stem cell-derived systems can be useful platforms to reveal novel functions of the apoptotic machinery in neural development.

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“…First samples were taken at day 3 (D3) of protocol, one day after medium change towards NMM with Dorsomorphin and SB431542. Second samples were taken at day 9 (D9), before rosette structures were cut, reminiscent to the early stage of secondary neurulation [26] followed by third sample collection at day 15 (D15), after rosettes were manually picked, comparable to complete neural tube formation state. Finally, fourth samples were taken at day 22 (D22), after first passage was performed and cells were recovered.…”

Section: Sample Collectionmentioning

confidence: 99%

Quantitative proteomic analysis shows alterations in patient Rett syndrome iPSC cultures at early neuronal progenitor stages

Varderidou-Minasian

Hinz

Hagemans

et al. 2020

Preprint

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BackgroundRett syndrome (RTT) is a progressive neurodevelopmental disease that is characterized by abnormalities in cognitive, social and motor skills. RTT is often caused by mutations in the X-linked gene encoding methyl-CpG binding protein 2 (MeCP2). The mechanism by which impaired MeCP2 induces the pathological abnormalities in the brain is not understood. Both patients and mouse models have shown abnormalities at molecular and cellular level before typical RTT-associated symptoms appear. This implies that underlying mechanisms are already affected during neurodevelopmental stages.MethodsTo understand the molecular mechanisms involved in disease onset, we used an RTT patient induced pluripotent stem cell (iPSC)-based model with isogenic controls and performed time-series of proteomic analysis using in-depth high-resolution quantitative mass spectrometry during early stages of neuronal development. ResultsWe provide mass spectrometry-based quantitative proteomic data, depth of about 7000 proteins, at neuronal progenitor developmental stages of RTT patient cells and isogenic controls. Our data gives evidence of proteomic alteration at early neurodevelopmental stages, suggesting alterations long before the phase that symptoms of RTT syndrome become apparent. We found changes in proteins involved in pathway associated with RTT phenotypes, including dendrite morphology and synaptogenesis. Differential expression increased from early to late neural stem cell phases, although proteins involved in immunity, metabolic processes and calcium signaling were affected throughout all stages analyzed. LimitationsThe limitation of our study is the number of biological replicates. As the aim of our study was to investigate a large number of proteins, only a limited amount of biological replicates were suitable for inclusions without reducing the number of target proteins. Therefore, larger sample sizes derived from RTT patients will be needed to validate results. ConclusionsOur results provide a valuable resource of proteins to study potential targets for early treatment of RTT symptoms. We found consistent and time-point specific alterations during early neuronal differentiation in RTT cultures. Insight into altered protein levels can help development of new biomarkers and therapeutic approaches in RTT syndrome. Therefore, we hope that our results give awareness of the early pre-natal onset of RTT, providing new insights to explore early diagnosis and treatment.

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Differentiation of neural rosettes from human pluripotent stem cells in vitro is sequentially regulated on a molecular level and accomplished by the mechanism reminiscent of secondary neurulation

Cited by 59 publications

References 45 publications

Neural Stem Cells and Methods for Their Generation From Induced Pluripotent Stem Cells in vitro

Neural Stem Cells and Methods for Their Generation From Induced Pluripotent Stem Cells in vitro

Modeling the function of BAX and BAK in early human brain development using iPSC-derived systems

Quantitative proteomic analysis shows alterations in patient Rett syndrome iPSC cultures at early neuronal progenitor stages

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