scholarly journals Human IPSC-Derived Model to Study Myelin Disruption

2021 ◽  
Vol 22 (17) ◽  
pp. 9473
Author(s):  
Megan Chesnut ◽  
Hélène Paschoud ◽  
Cendrine Repond ◽  
Lena Smirnova ◽  
Thomas Hartung ◽  
...  
Keyword(s):  
Proteolipid Protein ◽  
In Vitro Models ◽  
Negative Control ◽  
Reference Compound ◽  
Myelinated Axons ◽  
3D Cell Cultures ◽  
The Central Nervous System ◽  
Brain Organoid ◽  
Human Ipsc

Myelin is of vital importance to the central nervous system and its disruption is related to a large number of both neurodevelopmental and neurodegenerative diseases. The differences observed between human and rodent oligodendrocytes make animals inadequate for modeling these diseases. Although developing human in vitro models for oligodendrocytes and myelinated axons has been a great challenge, 3D cell cultures derived from iPSC are now available and able to partially reproduce the myelination process. We have previously developed a human iPSC-derived 3D brain organoid model (also called BrainSpheres) that contains a high percentage of myelinated axons and is highly reproducible. Here, we have further refined this technology by applying multiple readouts to study myelination disruption. Myelin was assessed by quantifying immunostaining/confocal microscopy of co-localized myelin basic protein (MBP) with neurofilament proteins as well as proteolipid protein 1 (PLP1). Levels of PLP1 were also assessed by Western blot. We identified compounds capable of inducing developmental neurotoxicity by disrupting myelin in a systematic review to evaluate the relevance of our BrainSphere model for the study of the myelination/demyelination processes. Results demonstrated that the positive reference compound (cuprizone) and two of the three potential myelin disruptors tested (Bisphenol A, Tris(1,3-dichloro-2-propyl) phosphate, but not methyl mercury) decreased myelination, while ibuprofen (negative control) had no effect. Here, we define a methodology that allows quantification of myelin disruption and provides reference compounds for chemical-induced myelin disruption.

scholarly journals Effects of Platelet-Rich Plasma on Cellular Populations of the Central Nervous System: The Influence of Donor Age

2021 ◽  
Vol 22 (4) ◽  
pp. 1725
Author(s):  
Diego Delgado ◽  
Ane Miren Bilbao ◽  
Maider Beitia ◽  
Ane Garate ◽  
Pello Sánchez ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Cellular Response ◽  
Platelet Rich Plasma ◽  
Biological Response ◽  
In Vitro Models ◽  
Molecular Composition ◽  
Cellular Models ◽  
The Central Nervous System

Platelet-rich plasma (PRP) is a biologic therapy that promotes healing responses across multiple medical fields, including the central nervous system (CNS). The efficacy of this therapy depends on several factors such as the donor’s health status and age. This work aims to prove the effect of PRP on cellular models of the CNS, considering the differences between PRP from young and elderly donors. Two different PRP pools were prepared from donors 65–85 and 20–25 years old. The cellular and molecular composition of both PRPs were analyzed. Subsequently, the cellular response was evaluated in CNS in vitro models, studying proliferation, neurogenesis, synaptogenesis, and inflammation. While no differences in the cellular composition of PRPs were found, the molecular composition of the Young PRP showed lower levels of inflammatory molecules such as CCL-11, as well as the presence of other factors not found in Aged PRP (GDF-11). Although both PRPs had effects in terms of reducing neural progenitor cell apoptosis, stabilizing neuronal synapses, and decreasing inflammation in the microglia, the effect of the Young PRP was more pronounced. In conclusion, the molecular composition of the PRP, conditioned by the age of the donors, affects the magnitude of the biological response.

scholarly journals Emergence of three myelin proteins in oligodendrocytes cultured without neurons.

1986 ◽  
Vol 102 (2) ◽  
pp. 384-392 ◽  
Author(s):  
M Dubois-Dalcq ◽  
T Behar ◽  
L Hudson ◽  
R A Lazzarini
Keyword(s):  
Optic Nerve ◽  
Proteolipid Protein ◽  
Defined Medium ◽  
Myelin Proteins ◽  
Specific Marker ◽  
The Central Nervous System ◽  
Double Label ◽  
Myelin Associated Glycoprotein

Oligodendrocytes, the myelin-forming cells of the central nervous system, were cultured from newborn rat brain and optic nerve to allow us to analyze whether two transmembranous myelin proteins, myelin-associated glycoprotein (MAG) and proteolipid protein (PLP), were expressed together with myelin basic protein (MBP) in defined medium with low serum and in the absence of neurons. Using double label immunofluorescence, we investigated when and where these three myelin proteins appeared in cells expressing galactocerebroside (GC), a specific marker for the oligodendrocyte membrane. We found that a proportion of oligodendrocytes derived from brain and optic nerve invariably express MBP, MAG, and PLP about a week after the emergence of GC, which occurs around birth. In brain-derived oligodendrocytes, MBP and MAG first emerge between the fifth and the seventh day after birth, followed by PLP 1 to 2 d later. All three proteins were confined to the cell body at that time, although an extensive network of GC positive processes had already developed. Each protein shows a specific cytoplasmic localization: diffuse for MBP, mostly perinuclear for MAG, and particulate for PLP. Interestingly, MAG, which may be involved in glial-axon interactions, is the first myelin protein detected in the processes at approximately 10 d after birth. MBP and PLP are only seen in these locations after 15 d. All GC-positive cells express the three myelin proteins by day 19. Simultaneously, numerous membrane and myelin whorls accumulate along the oligodendrocyte surface. The sequential emergence, cytoplasmic location, and peak of expression of these three myelin proteins in vitro follow a pattern similar to that described in vivo and, therefore, are independent of continuous neuronal influences. Such cultures provide a convenient system to study factors regulating expression of myelin proteins.

scholarly journals Dynamic 3D On-Chip BBB Model Design, Development, and Applications in Neurological Diseases

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 3183
Author(s):  
Xingchi Chen ◽  
Chang Liu ◽  
Laureana Muok ◽  
Changchun Zeng ◽  
Yan Li
Keyword(s):  
Targeted Delivery ◽  
Neurological Diseases ◽  
In Vitro Models ◽  
Normal Function ◽  
Human Stem Cells ◽  
Design Development ◽  
High Throughput Drug Screening ◽  
The Central Nervous System ◽  
On Chip

The blood–brain barrier (BBB) is a vital structure for maintaining homeostasis between the blood and the brain in the central nervous system (CNS). Biomolecule exchange, ion balance, nutrition delivery, and toxic molecule prevention rely on the normal function of the BBB. The dysfunction and the dysregulation of the BBB leads to the progression of neurological disorders and neurodegeneration. Therefore, in vitro BBB models can facilitate the investigation for proper therapies. As the demand increases, it is urgent to develop a more efficient and more physiologically relevant BBB model. In this review, the development of the microfluidics platform for the applications in neuroscience is summarized. This article focuses on the characterizations of in vitro BBB models derived from human stem cells and discusses the development of various types of in vitro models. The microfluidics-based system and BBB-on-chip models should provide a better platform for high-throughput drug-screening and targeted delivery.

scholarly journals In vitro models of HIV-1 infection of the central nervous system

2020 ◽  
Vol 32 ◽  
pp. 5-11
Author(s):  
Celeste Faia ◽  
Karlie Plaisance-Bonstaff ◽  
Francesca Peruzzi
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
In Vitro Models ◽  
The Central Nervous System ◽  
Hiv 1

scholarly journals Mass Generation, Neuron Labelling and 3D Imaging of Minibrains

2020 ◽  
Author(s):  
Subashika Govindan ◽  
Laura Batti ◽  
Samira F Osterop ◽  
Luc Stoppini ◽  
Adrien Roux
Keyword(s):  
Large Scale ◽  
3D Imaging ◽  
Cell Types ◽  
Projection Neurons ◽  
Neuron Morphology ◽  
Mass Generation ◽  
One Step ◽  
Brain Organoid ◽  
Human Ipsc

AbstractMinibrain is a spherical in vitro 3D brain organoid model, composed of a mixed population of neurons and glial cells, generated from human iPSC derived neural stem cells. Despite the advances in human brain organoid models, there is a lack of labelling and imaging methodologies to characterize these models. In this study, we present a step-by-step methodology to generate human minibrain nurseries and novel strategies to subsequently label projection neurons, perform immunohistochemistry and 3D imaging of the minibrains at large multiplexable scales. To visualize projection neurons, we adapt viral transduction and to visualize the organization of cell types we implement immunohistochemistry. To facilitate 3D imaging of minibrains, we present here pipelines and accessories for one step mounting and clearing suitable for confocal microscopy. The pipelines are specifically designed in such a way that the assays can be multiplexed with ease for large-scale screenings using minibrains. Using the pipeline, we present i. dendrite morphometric properties obtained from 3D neuron morphology reconstructions and ii. distribution and quantification of cell types in 3D across whole mount organoids.

The molecular and cellular defects underlying Pelizaeus–Merzbacher disease

2008 ◽  
Vol 10 ◽  
Author(s):  
Karen J. Woodward
Keyword(s):  
Current Knowledge ◽  
Point Mutations ◽  
Proteolipid Protein ◽  
Major Protein ◽  
Cellular Mechanisms ◽  
Underlying Causes ◽  
The Central Nervous System ◽  
Genomic Disorders ◽  
Pelizaeus Merzbacher Disease

Pelizaeus–Merzbacher disease (PMD) is a recessive X-linked dysmyelinating disorder of the central nervous system (CNS). The most frequent cause of PMD is a genomic duplication of chromosome Xq22 including the region encoding the dosage-sensitive proteolipid protein 1 (PLP1) gene. The PLP1 duplications are heterogeneous in size, unlike duplications causing many other genomic disorders, and arise by a distinct molecular mechanism. Other causes of PMD include PLP1 deletions, triplications and point mutations. Mutations in the PLP1 gene can also give rise to spastic paraplegia type 2 (SPG2), an allelic form of the disease. Thus, there is a spectrum of CNS disorder from mild SPG2 to severe connatal PMD. PLP1 encodes a major protein in CNS myelin and is abundantly expressed in oligodendrocytes, the myelinating cells of the CNS. Significant advances in our understanding of PMD have been achieved by investigating mutant PLP1 in PMD patients, animal models and in vitro studies. How the different PLP1 mutations and dosage effects give rise to PMD is being revealed. Interestingly, the underlying causes of pathogenesis are distinct for each of the different genetic abnormalities. This article reviews the genetics of PMD and summarises the current knowledge of causative molecular and cellular mechanisms.

Neural In Vitro Models for Studying Substances Acting on the Central Nervous System

2020 ◽  
Author(s):  
Ellen Fritsche ◽  
Julia Tigges ◽  
Julia Hartmann ◽  
Julia Kapr ◽  
Melania Maria Serafini ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
In Vitro Models ◽  
The Central Nervous System

scholarly journals Induced Pluripotent Stem Cell Models to Enable In Vitro Models for Screening in the Central Nervous System

2015 ◽  
Vol 24 (16) ◽  
pp. 1852-1864 ◽  
Author(s):  
Joshua G. Hunsberger ◽  
Anastasia G. Efthymiou ◽  
Nasir Malik ◽  
Mamta Behl ◽  
Ivy L. Mead ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
In Vitro Models ◽  
Cell Models ◽  
The Central Nervous System ◽  
Induced Pluripotent ◽  
Stem Cell Models

scholarly journals Senescence and associated blood–brain barrier alterations in vitro

2021 ◽  
Author(s):  
Ellaine Salvador ◽  
Malgorzata Burek ◽  
Mario Löhr ◽  
Michiaki Nagai ◽  
Carsten Hagemann ◽  
...  
Keyword(s):  
Blood Brain Barrier ◽  
Intercellular Junctions ◽  
In Vitro Models ◽  
Brain Barrier ◽  
Initial Attempt ◽  
Blood Brain ◽  
The Central Nervous System ◽  
Vitro Model ◽  
The Relationship

AbstractProgressive deterioration of the central nervous system (CNS) is commonly associated with aging. An important component of the neurovasculature is the blood–brain barrier (BBB), majorly made up of endothelial cells joined together by intercellular junctions. The relationship between senescence and changes in the BBB has not yet been thoroughly explored. Moreover, the lack of in vitro models for the study of the mechanisms involved in those changes impede further and more in-depth investigations in the field. For this reason, we herein present an in vitro model of the senescent BBB and an initial attempt to identify senescence-associated alterations within.

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