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Influence of glucocorticoids on microglia development
December 2022
University College London Institute of Neurology, London, United Kingdom
Prenatal exposure to glucocorticoids (GC) results in a higher risk of psychiatric and neurodevelopmental disorders in later life. In this study, induced pluripotent stem cells (iPSCs) were generated from fibroblasts derived from patients and healthy individuals. iPSCs were exposed to GC during early differentiation into microglia, and it was examined how this exposure affected the microglial phenotype using RNA-seq analyses and functional assays.
The in vitro model allows the investigation of the responses of microglia in the pathogenesis of prenatal GC- exposure-associated disorders such as schizophrenia, attention-deficit hyperactivity disorder and autism spectrum disorder.
Human myeloid progenitor glucocorticoid receptor activation causes genomic instability, type 1 IFN- response pathway activation and senescence in differentiated microglia; an early life stress model
Jennifer M. Pocock, Thomas M. Piers
Added on: 10-26-2023
COVID-19-related immune signatures and neurological sequelae
November 2022
University Hospital Basel, Basel, Switzerland
COVID-19 is associated with acute and long-term neurological dysfunction, but the pathophysiological mechanisms resulting in central nervous system involvement remain unclear.
To address this issue, a cross-sectional clinical study was performed, including clinical and imaging data and corresponding multidimensional characterization of immune mediators in the cerebrospinal fluid (CSF) and plasma of patients belonging to different Neuro-COVID severity classes.
40 COVID-19 patients, 25 healthy controls and 25 patients with neurological inflammation of other causes were included in the study and the findings obtained from clinical sample analysis were correlated with brain imaging.
The most prominent signs of severe Neuro-COVID are blood-brain barrier impairment, elevated microglia activation markers and a polyclonal B cell response targeting self-antigens and non-self-antigens. Specific CSF and plasma alterations were identified, which provide insights into the pathomechanism underlying COVID-19-related neurological sequelae. Moreover, several potentially actionable targets to prevent or intervene with the neurological consequences of SARS-CoV-2 infection were identified.
Severe Neuro-COVID is associated with peripheral immune signatures, autoimmunity and neurodegeneration: a prospective cross-sectional study
Gregor Hutter
Added on: 11-21-2022
Brain-model for Parkinson´s disease
CompanyOctober 2021
Emulate Inc., Boston, USA(1)
Harvard Medical School, Boston, USA(2)
Harvard Medical School, Boston, USA(2)
A human brain-chip representative of the substantia nigra area of the brain, the area predominantly affected in Parkinson´s disease, was developed. It contains human dopaminergic neurons, astrocytes, microglia, pericytes, and microvascular brain endothelial cells, cultured under fluid flow, thereby recreating the vascular-neuronal interface. A model of neurodegenerative diseases characterized by the accumulation of aggregated α-synuclein (synucleinopathies) was created. Therefore, human α-synuclein pre-formed fibrils were introduced within the brain channel of the chip. It was demonstrated that the model replicates pathological key aspects observed in human Parkinson´s disease, including accumulation of α-Synuclein, mitochondrial dysfunction, neuroinflammation and compromised blood-brain barrier function. The human α-synuclein fibril-induced disease model could provide a model for recapitulating complex pathophysiological features of Parkinson´s disease, may enable research of the dynamics of cell-cell interactions in human synucleinopathies, and serve as a testing platform for target identification and validation of novel therapeutics.
Modeling alpha-synuclein pathology in a human brain-chip to assess blood-brain barrier disruption
Iosif Pediaditakis(1), Katia Karalis(2)
Added on: 03-31-2022
Mini brains to study motor neuron disease and frontotemporal dementia
October 2021
University of Cambridge, Cambridge, United Kingdom
Amyotrophic lateral sclerosis overlapping with frontotemporal dementia (ALS/FTD) is a fatal and currently untreatable disease characterized by rapid cognitive decline and paralysis. Elucidating the initial cellular pathologies is central to developing therapeutic targets, but it is not possible to obtain samples from presymptomatic patients. Here, the researchers develop a long-term human cortical organoid (CO) model that accurately recapitulates the early molecular pathology of ALS/FTD. COs were grown from iPSCs derived from patients with ALS/FTD carrying the C9ORF72 mutation (C9 ALI-COs). This mutation is useful for modelling ALS/FTD as it causes a variety of pathologies in both sporadic and inherited forms of the disease. Using ALI-CO discs cultured from the organoids, ALI-COs are shown to form consistent microarchitecture and disease-relevant phenotypes. Furthermore, it was found that C9 ALI-COs, although lacking microglia and vasculature, exhibit astroglia- and neuron-specific perturbations. These results demonstrate that hiPSC-derived ALI-COs provide a reproducible platform with the necessary longevity and maturity to study ALS/FTD, thereby revealing early and targeted cellular vulnerabilities relevant to presymptomatic clinical stages.
Human ALS/FTD brain organoid slice cultures display distinct early astrocyte and targetable neuronal pathology
András Lakatos, Gabriel Balmus
Added on: 11-09-2021
Human brain organoids develop eyes
2021
Heinrich-Heine-Universität, Dusseldorf, Germany
Here, iPSC technology is used to grow mini-brains in order to study inherited eye diseases. With cells from four iPSC donors, brain organoids were grown and on day 20, retinoic acid was added. After 30 days of cultivation, the organoids assembled optical vesicles, which developed within 60 days to visible structures. The optic vesicle-containing brain organoids (OVB-organoids) show various structures which are typical for the eye like primitive corneal epithelial and lens-like cells, retinal pigment epithelia, retinal progenitor cells, axon-like projections, and electrically active neuronal networks. They as well display synapsin-1, CTIP-positive myelinated cortical neurons, and microglia. The organoids even show photosensitive activity when triggered by various light intensities, which could be reset by transient photobleaching. The OVB-organoids possess the ability to self-organize forebrain-associated primitive sensory structures and thus can be used as a model to study eye development and underlying causes of eye diseases.
Human brain organoids assemble functionally integrated bilateral optic vesicles
Jay Gopalakrishnan
Added on: 09-09-2021
Development of microglial-containing human brain organoids
2021
Rutgers University, Piscataway, USA
Microglia is an essential actor in brain physiology. In recent years, human pluripotent stem cells are surging as a reliable tool to generate specific cell types to incorporate in in vitro models. Nonetheless, there are still no optimised protocols to properly recapitulate microglia differentiation in in vivo brain development. Here, human pluripotent stem cells-derived neural and macrophage progenitor cells were used to generate region-specific brain organoids that contain microglia. The results showed that the generation of organoids had a controllable microglia ratio and that the microglial cells were physiologically functional. Moreover, Zika virus infection efficiently triggered microglial responses. Overall, the researchers establish a new protocol replicating in vivo neurodevelopment for the generation of organoids including microglia that could potentially be used to study microglial physiology and its role in neurological disorders.
Developing human pluripotent stem cell-based cerebral organoids with a controllable microglia ratio for modeling brain development and pathology
Peng Jiang, Ranjie Xu
Added on: 11-29-2021
COVID-19 can cause severe inflammation in the brain
2021
University Medical Center Freiburg, Freiburg, Germany(1)
University of Freiburg, Freiburg, Germany(2)
University of Freiburg, Freiburg, Germany(2)
The researchers interrogated the brain stem and olfactory bulb in brain samples of COVID-19 patients postmortem using imaging mass cytometry to understand the local immune response at a spatially resolved, high-dimensional single-cell level. They compared this immune map to non-COVID respiratory failure, multiple sclerosis and control patients. The findings show that a severe inflammatory response can develop in the central nervous system of COVID-19 patients involving different immune cells around the vascular system and in the brain tissue. This study identifies profound neuroinflammation with activation of innate and adaptive immune cells as correlates of COVID-19 neuropathology, with implications for potential therapeutic strategies.
Deep spatial profiling of human COVID-19 brains reveals neuroinflammation with distinct microanatomical microglia-T cell interactions
Bertram Bengsch(1), Marco Prinz(2)
Added on: 06-22-2021
iPSC-derived microglia for research of Alzheimer's disease
Company 2021
F. Hoffmann-La Roche Ltd., Basel, Switzerland
Microglia are key in the homeostatic well-being of the brain and microglial dysfunction has been implicated in neurodegenerative disorders such as Alzheimer’s disease (AD). Due to the many limitations to study microglia in situ or isolated for large scale drug discovery applications, there is a high need to develop robust and scalable human cellular models of microglia.
Here, the optimization of a protocol to generate microglia from iPSCs in a monoculture condition is presented. The study explores whether these cells can serve as a model to study microglial function and gene expression in the context of TREM2 modulation. TREM2 is a risk gene for AD and an important regulator of microglia. As a major distinction from previously published co-culture methods, in iPSC-generated microglia from monoculture, an increased TREM2 mRNA expression was observed. The regulatory function of TREM2 in these cells was confirmed by comparing wild type with isogenic TREM2 knock-out iPSC microglia. The overall approach resulted in a workstream to generate human iPSC microglia by a directed and neuronal co-culture independent differentiation, resulting in distinct phenotypes for mechanistic studies in AD. This iPSC microglia protocol can now be applied to scale up the production of these cells, study certain AD-related disease settings, and perform compound screening and mechanistic experiments in drug development.
Alzheimer’s risk gene TREM2 determines functional properties of new type of human iPSC-derived microglia
Simon Gutbier, Markus Britschgi
Added on: 05-11-2021
Brain organoids for HIV research
2020
University of Pittsburgh, Pittsburgh, USA
The infection with HIV-1 can result in HIV-associated neurocognitive disorder (HAND), which is characterized by neuroinflammation and neurodegeneration and results in cognitive impairment of affected patients. The aim of this study was to develop a brain-representative in-vitro system to investigate the molecular events underlying HIV neuropathogenesis.
Therefore, a three-dimensional (3D) human brain organoid (hBORG) model containing major cell types important for HIV-1 neuropathogenesis; in particular neurons and astrocytes, was developed. The ability of these hBORGs to support HIV-1 infection as well as to recapitulate the hallmarks of central nervous system (CNS) pathology seen in HIV-1 patients was investigated by incorporating HIV-infected primary microglia. Incorporation of HIV-infected microglia resulted in inflammatory response and induced damage to neurons and astrocytes, which represents major hallmark features seen in the CNS of HIV-1 infected individuals.
This model offers great promise for basic understanding of how HIV-1 infection alters the CNS and induces pathological changes, paving the way for discovery of biomarkers and new therapeutic targets.
Modeling HIV‑1 neuropathogenesis using three‑dimensional human brain organoids (hBORGs) with HIV‑1 infected microglia
Shilpa Sant, Velpandi Ayyavoo
Added on: 04-03-2023
3D organoid made of human cells to model the blood-brain barrier and screen drugs
2020
Wake Forest School of Medicine, Winston-Salem, USA
The blood-brain barrier (BBB) comprises a complex arrangement of various cell types which constitute a physical and enzymatic barrier between the brain and the bloodstream. BBB is essential for maintaining the brain's homeostasis. During ischemic stroke, BBB breakdown leads to oedema and haemorrhage which cause major brain damage. The lack of an agreed ischemic stroke model limits the search for molecular therapies. In the present study, the researchers aimed at developing a six cell-type neurovascular unit human organoid model for use in neurotoxicity screening and disease modelling. Human brain microvascular endothelial cells, pericytes, astrocytes, oligodendrocytes, microglia and neurons were obtained from primary material or differentiated from induced pluripotent stem cells. The six cell types were assembled into a 3D organoid in vitro that was then cultured in hypoxia conditions to mimic stroke. The researchers measured changes in expression levels of proteins critical in BBB maintenance/function. The secretion and effect of inflammatory mediators were also assessed. The changes due to hypoxia could be lowered using drugs known to act on hypoxia stress and inflammation, hence validating the model. The study concludes that such neurovascular spheroid is a suitable model for mimicking cerebral pathology, such as hypoxia, that will allow for in vitro testing and the development of novel therapies for diseases of the central nervous system.
Multicellular 3D neurovascular unit model for assessing hypoxia and neuroinflammation induced blood-brain barrier dysfunction
Goodwell Nzou
Added on: 12-20-2021
Neuronal organoids with astrocytes for toxicological studies
2020
Universität Konstanz, Konstanz, Germany
A fast and robust method to generate 3D cultured human dopaminergic neurons (LUHMES) for toxicity testing and long-term culture is presented. A special plating step allows the generation of neurite networks with defined 2D orientation, while all cell bodies (somata) remain in a 3D, dome-like structure. These cultures, named 2.5 dimensional, offer new approaches to quantify toxic effects on organoids by standard technology and high throughput. For instance, the system reacts to the parkinsonian model toxicants MPP+, rotenone, MG-132 and the ferroptosis-inducer erastin. Moreover, stable incorporation of human stem cell-derived astrocytes or microglia is possible. Added astrocytes stabilize the postmitotic state of the LUHMES neurons and thereby allow the formation of a stable microphysiological system. Neuroprotection against the proteasome inhibitor MG-132 and the ferroptosis-inducer erastin by such glia is observed. This exemplifies the crucial protective role of astrocytes in neurodegeneration. The modularity of the system is further employed to incorporate microglia together with astrocytes into the organoids. Such ratio-defined, three cell type-based organoids will allow new approaches to study human pathophysiology and toxicology of the nervous system.
Incorporation of stem cell-derived astrocytes into neuronal organoids to allow neuro-glial interactions in toxicological studies
Marcel Leist
Added on: 07-07-2020
3D-Brainspheres as model for nanoparticle-associated substances
2019
Center for Alternatives to Animal Testing (CAAT), Johns Hopkins Bloomberg School of Public Health, Baltimore, USA
Nanoparticles (NP) can cross the blood-brain-barrier (BBB) and accumulate in different areas of the central nervous system (CNS), thus are potential tools to carry drugs and treat brain disorders. The effects of different NP were studied in human LUHMES cell line (3D LUHMES) and human iPSC-derived brain spheroids (BrainSpheres). The simpler, single-cell model was more sensitive to the toxic effects, in line with the lack of glia support to neurons. In BrainSpheres the NP, especially modified with specific ligands, can cross the BBB and show higher levels due to the presence of glial cells whose function is to facilitate uptake. Nevertheless, decrease of viability or morphological alterations was not observed, maybe due to activation of cell survival programs.
Therefore, 3D brain spheroid models incorporating microglia are well suited to comparatively characterize NP neurotoxicity. The use of multiple models, which encompass simplicity and physiological relevance, serves as a tool for more NP drug-delivery focused research.
Suitability of 3D human brain spheroid models to distinguish toxic effects of gold and poly-lactic acid nanoparticles to assess biocompatibility for brain drug delivery
Lena Smirnova
Added on: 08-28-2020
Human brain model to study Dengue and Zika virus
December 2018
Center for Alternatives to Animal Testing (CAAT), Johns Hopkins Bloomberg School of Public Health, Baltimore, USA
It is shown that brain organoids containing microglia cells in addition to astrocytes, oligodendrocytes and neuronal populations possess physiologically relevant features. Without these cells, the organoids do not show inflammatory reactions to e.g. lipopolysaccharides (LPS) or flaviviruses. By co-cultivation with microglia cells, the gene expression of inflammatory cytokines is induced in the brain organoids. After infection with Zika and Dengue viruses interleukins, tumor necrosis factor alpha (TNF-α) and chemokines are also induced.
This shows that the model is physiologically relevant and has potential applications in research on infectious disease and host-pathogen interactions.
Microglia Increase Inflammatory Responses in iPSC-Derived Human BrainSpheres
David Pamies
Added on: 08-28-2020
3D multicellular microfluidic platform to model neurodegenerative diseases
2018
University of North Carolina at Charlotte, Charlotte, USA
Mechanisms of Alzheimer's disease pathology remain poorly understood. In this study, the researchers develop a 3D multicellular model that integrates human neuronal cells, astrocytes and microglia in a microfluidic platform. This system recapitulated the key hallmarks of the disease that involved the different cell types present in the model, allowing the study of intercellular interactions relevant for the pathophysiology of Alzheimer's disease. In conclusion, this new model can help to develop better human brain models for mechanistic studies of neurodegenerative diseases that involve neuro-glial interactions and improve drug discovery.
A 3D human triculture system modeling neurodegeneration and neuroinflammation in Alzheimer’s disease
Hansang Cho
Added on: 08-04-2021
Multicellular organoids with blood-brain barrier for neurotoxicity screening
2018
Wake Forest School of Medicine, Winston-Salem, USA
The development of a human neurovascular organoid model is reported. The model contains the six constituent cell types found within the brain cortex: human endothelial cells, pericytes, astrocytes, microglia, oligodendrocytes and neurons, with endothelial cells enclosing the brain parenchymal cells. Cells derived from induced pluripotent stem cell (iPSC) sources were utilized to establish potential patient-specific and disease applications. It was shown that the model can be used in toxicity assessment studies for molecules that have the potential to cross or open the blood-brain barrier. Neurotoxicity screening was assessed and a model of the blood-brain barrier during clinical ischemia was established showing physiologic responses under hypoxic conditions. Taken together, the multicellular organoid model forms a functional blood-brain barrier and may be useful not only in drug discovery of novel therapeutics but also to evaluate the ability of drug candidates to cross the blood-brain barrier.
Human cortex spheroid with a functional blood brain barrier for high-throughput neurotoxicity screening and disease modeling
Doodwell Nzou
Added on: 03-16-2022
Protocol to culture human microglia
October 2016
Northwestern University, Chicago, USA
Microglia are the resident macrophages of the central nervous system. They are immune cells and have a wide range of physiological functions. Our current knowledge of this cell type relies mainly on rodent models, but differences in biological features with human samples have shown the need to obtain reliable techniques to use human microglia in in vitro studies. The main limitation for this is that currently there is no protocol that offers the opportunity to maintain human microglia in culture for a long time and for a high number of passages. To solve this, in this study a new protocol is developed based on the use of several growth factors that overcomes this limitation. The cells maintained their features and functions even in high passage numbers. Also, cultured human microglia was responsive to different activators. Additionally, microglia from Alzheimer's disease brains showed the same characteristics as from normal brains. The researchers propose a new method to be able to reliably culture microglia for long-term and high passages, which will give the opportunity to investigators to extend the mechanistic studies in human microglia from different patient profiles.
Postmortem adult human microglia proliferate in culture to high passage and maintain their response to amyloid-β
Changiz Geula
Added on: 08-23-2021
Neural constructs for predicting neural toxicity
2015
Morgridge Institute for Research, Madison, USA
Human pluripotent stem cell-based in vitro models that mirror human physiology have the potential to cost-effectively assess the developmental neurotoxicity of chemicals. Here, human embryonic stem (ES)-derived neural progenitor cells, endothelial cells, mesenchymal stem cells and microglia/macrophage progenitors were combined on synthetic hydrogels and cultured in a serum-free medium to model cellular interactions in the developing brain. The progenitor cells self-assembled into 3D neuronal constructs with distinct populations of neurons and glia, interconnected vascular networks and branching microglia. The replicate constructs were reproducible by RNA sequencing (RNA-Seq) and expressed genes for neurogenesis, vascular development and microglia. Using machine learning, a predictive model was built from these RNA-Seq for the neuronal constructs exposed to a training set of 60 toxic and non-toxic chemicals and then predicted in a blind trial with a set of 10 additional compounds. The model correctly classified 9 of the 10 additional chemicals. This combined strategy demonstrates the value of cell-based assays for predictive toxicology and should be useful for assessing the safety of both drugs and chemicals.
Human pluripotent stem cell-derived neural constructs for predicting neural toxicity
James A. Thomson
Added on: 11-09-2021
New human microglial marker
2015
Meiji Pharmaceutical University, Tokyo, Japan
Microglia are the resident macrophages of the central nervous system. In recent years, they have been described to have an important role in several neurological disorders. However, it is still difficult to discriminate resident microglial cells from infiltrating macrophages and this interferes with the study of human microglia. In this study, the researchers try to find a new marker that is exclusively and consistently expressed in human microglia. After a first comparative analysis in microglial transcriptome datasets, TMEM119 is identified as a candidate marker, which is found to be consistently expressed in immortalised human microglia in different conditions. This marker is also described to be a better discriminator from infiltrating macrophages that express currently used microglial markers. Finally, there were no differences in the expression of TMEM119 in different subsets of microglia and between control and diseased human brains, clarifying that this can be a reliable marker to be used to study microglia in different conditions. These results offer new possibilities to better isolate and study human microglia and show the possibility of using human models to improve the translational value of glial biology studies.
TMEM119 marks a subset of microglia in the human brain
Jun-ichi Satoh
Added on: 08-16-2021
Development of primary mixed human brain cultures
2014
Indiana University School of Medicine, Indianapolis, USA
Primary cultures are a basic tool in neuroscience. However, most of the time these cultures are done with rodent brains, associated with several limitations in the translational potential of the model. In this study, human fetal brains are used to produce mixed primary brain cultures. The results show characteristic features in human cultures not present in animal-based models. With the proper methods, it is possible to maintain brain cells for a long time in culture, producing different types of neurons and glia. Additionally, consistent production of amyloid-beta, tau and other characteristic markers of Alzheimer's disease is also described. Basically, the study presents a new human model with a detailed characterization that can be a powerful tool to enhance the translationality of neuroscience studies and help to better understand the mechanisms of human brain disorders, leading to new therapeutic approaches.
Human primary mixed brain cultures: preparation, differentiation, characterization and application to neuroscience research
Debomoy K Lahiri
Added on: 08-22-2021
Development of 3D brain organoids to study neuronal plasticity
Companymyriamed GmbH, Goettingen, Germany
The biotechnologically engineered neuronal organoids (BENOs) from myriamed consist of excitatory (glutamatergic) and inhibitory (GABAergic) neurons, as well as supporting glial cells (astrocytes, oligodendrocytes). The stem cell-derived cells are cultivated in a collagen hydrogel in which they form highly interconnected, three-dimensional neural networks. Studies have shown that BENOs recapitulate important steps in fetal development of the human brain, such as the appearance of giant depolarizing potentials (GDPs). After more than 40 days in culture, a reduction in GDPs and a GABA polarity switch were observed, indicating progressive maturation of the mini brain models. After two months, an accelerated development and an increased occurrence of neuronal networks, as well as indications of long-term potentiation, could be observed. BENOs therefore enable a deeper investigation of the neuronal plasticity of the brain, as well as a modelling of neuronal diseases. The brain organoids are suitable for a variety of (pre-)clinical studies and can help to expand the understanding of the development and maturation processes of the brain and to improve drug development.
myrTissue-Brain
contact@myriamed.com
Added on: 08-16-2023
hiPSC-based neurons for modelling neurological diseases and assessing neurotoxicities
CompanyFUJIFILM Cellular Dynamics Inc., Madison, USA
The nerve cells from FUJIFILM Cellular Dynamics Inc. (FCDI) obtained from human induced pluripotent stem cells (hiPSCs) enable the development of complex, three-dimensional mini-brain models for researching neurological processes and diseases. The iCell kits include a range of different cell types such as: astrocytes, microglia, motor neurons, retinal pigment epithelial cells, various neurotransmitter-associated nerve cells (dopa, GABA and gluta neurons), as well as a specially developed cell kit for modeling the blood-brain barrier. The iCell Blood-Brain Barrier Isogen Kit is of particular relevance for drug research as it enables the evaluation of drugs to be tested with regard to possible neurotoxicities, as well as an assessment of their permeability and their possible influence on the barrier function of the blood-brain barrier. The cell kits are suitable for modelling different diseases of the central nervous system; ranging from neurodegenerative diseases, such as Parkinson's or Alzheimer's, to neurodevelopmental diseases, such as autism spectrum disorders (ASD). FCDI offers the cryopreserved nerve cells in a healthy state and with pre-programmed diseases. The neural iCell kits are therefore suitable for a variety of (pre-)clinical applications and studies and can help to expand the understanding of neuropathological processes and improve the drug therapy of neurological diseases.
iCell iPSC neurons
www.fujifilmcdi.com
Added on: 09-21-2023
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