Brain tumor invasion model system using organotypic brain-slice culture as an alternative to in vivo model

2002 ◽  
Vol 128 (9) ◽  
pp. 469-476 ◽  
Author(s):  
Shin Jung ◽  
Hyun-Woo Kim ◽  
Je-Hyuk Lee ◽  
Sam-Suk Kang ◽  
Hyang-Hwa Rhu ◽  
...  
Keyword(s):  
Brain Tumor ◽  
Tumor Invasion ◽  
Brain Slice ◽  
Model System ◽  
In Vivo Model ◽  
Slice Culture ◽  
Invasion Model ◽  
Organotypic Brain Slice ◽  
Brain Slice Culture

scholarly journals Modeling α-Synucleinopathy in Organotypic Brain Slice Culture with Preformed α-Synuclein Amyloid Fibrils

2020 ◽  
Vol 10 (4) ◽  
pp. 1397-1410
Author(s):  
Amandine Roux ◽  
Xinhe Wang ◽  
Katelyn Becker ◽  
Jiyan Ma
Keyword(s):  
Brain Slice ◽  
Amyloid Fibrils ◽  
Three Dimensional ◽  
Brain Slices ◽  
Alpha Synuclein ◽  
Slice Culture ◽  
Suitable Model ◽  
Organotypic Brain Slice ◽  
Brain Slice Culture

Background: Synucleinopathy is a group of neurodegenerative disorders characterized by neurodegeneration and accumulation of alpha-synuclein (α-syn) aggregates in various brain regions. The detailed mechanism of α-syn-caused neurotoxicity remains obscure, which is partly due to the lack of a suitable model that retains the in vivo three-dimensional cellular network and allows a convenient dissection of the neurotoxic pathways. Recent studies revealed that the pre-formed recombinant α-syn amyloid fibrils (PFFs) induce a robust accumulation of pathogenic α-syn species in cultured cells and animals. Objective: Our goal is to determine whether PFFs are able to induce the pathogenic α-syn accumulation and neurotoxicity in organotypic brain slice culture, an ex vivo system that retains the in vivo three-dimensional cell-cell connections. Methods/Results: Adding PFFs to cultured wild-type rat or mouse brain slices induced a time-dependent accumulation of pathogenic α-syn species, which was indicated by α-syn phosphorylated at serine 129 (pα-syn). The PFF-induced pα-syn was abolished in brain slices prepared from α-syn null mice, suggesting that the pα-syn is from the phosphorylation of endogenous α-syn. Human PFFs also induced pα-syn in brain slices prepared from mice expressing human α-syn on a mouse α-syn-null background. Furthermore, the synaptophysin immunoreactivity was inversely associated with pα-syn accumulation and an increase of neuronal loss was detected. Conclusion: PFF-treatment of brain slices is able to induce key pathological features of synucleinopathy: pα-syn accumulation and neurotoxicity. This model will be useful for investigating the neurotoxic mechanism and evaluating efficacy of therapeutic approaches.

scholarly journals Preparation of organotypic brain slice cultures for the study of Alzheimer’s disease

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 592
Author(s):  
Cara L. Croft ◽  
Wendy Noble
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Brain Slice ◽  
Synaptic Dysfunction ◽  
Slice Culture ◽  
Therapeutic Effects ◽  
Culture Model ◽  
Organotypic Brain Slice ◽  
Brain Slice Cultures

Alzheimer's disease, the most common cause of dementia, is a progressive neurodegenerative disorder characterised by amyloid-beta deposits in extracellular plaques, intracellular neurofibrillary tangles of aggregated tau, synaptic dysfunction and neuronal death. There are no cures for AD and current medications only alleviate some disease symptoms. Transgenic rodent models to study Alzheimer’s mimic features of human disease such as age-dependent accumulation of abnormal beta-amyloid and tau, synaptic dysfunction, cognitive deficits and neurodegeneration. These models have proven vital for improving our understanding of the molecular mechanisms underlying AD and for identifying promising therapeutic approaches. However, modelling neurodegenerative disease in animals commonly involves aging animals until they develop harmful phenotypes, often coupled with invasive procedures. In vivo studies are also resource, labour, time and cost intensive. We have developed a novel organotypic brain slice culture model to study Alzheimer’ disease which brings the potential of substantially reducing the number of rodents used in dementia research from an estimated 20,000 per year. We obtain 36 brain slices from each mouse pup, considerably reducing the numbers of animals required to investigate multiple stages of disease. This tractable model also allows the opportunity to modulate multiple pathways in tissues from a single animal. We believe that this model will most benefit dementia researchers in the academic and drug discovery sectors. We validated the slice culture model against aged mice, showing that the molecular phenotype closely mimics that displayed in vivo, albeit in an accelerated timescale. We showed beneficial outcomes following treatment of slices with agents previously shown to have therapeutic effects in vivo, and we also identified new mechanisms of action of other compounds. Thus, organotypic brain slice cultures from transgenic mouse models expressing Alzheimer’s disease-related genes may provide a valid and sensitive replacement for in vivo studies that do not involve behavioural analysis.

Invasion and Anti-Invasion Research of Glioma Cells in An Improved Model of Organotypic Brain Slice Culture

2015 ◽  
Vol 101 (4) ◽  
pp. 390-397 ◽  
Author(s):  
Bingcheng Ren ◽  
Shengping Yu ◽  
Cong Chen ◽  
Le Wang ◽  
Zhifeng Liu ◽  
...  
Keyword(s):  
Brain Slice ◽  
Glioma Cells ◽  
Slice Culture ◽  
Organotypic Brain Slice ◽  
Brain Slice Culture ◽  
Improved Model

scholarly journals Preparation of organotypic brain slice cultures for the study of Alzheimer’s disease

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 592 ◽  
Author(s):  
Cara L. Croft ◽  
Wendy Noble
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Brain Slice ◽  
Synaptic Dysfunction ◽  
Slice Culture ◽  
Therapeutic Effects ◽  
Culture Model ◽  
Organotypic Brain Slice ◽  
Brain Slice Cultures

Alzheimer's disease, the most common cause of dementia, is a progressive neurodegenerative disorder characterised by amyloid-beta deposits in extracellular plaques, intracellular neurofibrillary tangles of aggregated tau, synaptic dysfunction and neuronal death. Transgenic rodent models to study Alzheimer’s mimic features of human disease such as age-dependent accumulation of abnormal beta-amyloid and tau, synaptic dysfunction, cognitive deficits and neurodegeneration. These models have proven vital for improving our understanding of the molecular mechanisms underlying AD and for identifying promising therapeutic approaches. However, modelling neurodegenerative disease in animals commonly involves aging animals until they develop harmful phenotypes, often coupled with invasive procedures. We have developed a novel organotypic brain slice culture model to study Alzheimer’s disease using 3xTg-AD mice which brings the potential of substantially reducing the number of rodents used in dementia research from an estimated 20,000 per year. Using a McIllwain tissue chopper, we obtain 36 x 350 micron slices from each P8-P9 mouse pup for culture between 2 weeks and 6 months on semi-permeable 0.4 micron pore membranes, considerably reducing the numbers of animals required to investigate multiple stages of disease. This tractable model also allows the opportunity to modulate multiple pathways in tissues from a single animal. We believe that this model will most benefit dementia researchers in the academic and drug discovery sectors. We validated the slice culture model against aged mice, showing that the molecular phenotype closely mimics that displayed in vivo, albeit in an accelerated timescale. We showed beneficial outcomes following treatment of slices with agents previously shown to have therapeutic effects in vivo, and we also identified new mechanisms of action of other compounds. Thus, organotypic brain slice cultures from transgenic mouse models expressing Alzheimer’s disease-related genes may provide a valid and sensitive replacement for in vivo studies that do not involve behavioural analysis.

A novel method of organotypic brain slice culture using connexin-specific antisense oligodeoxynucleotides to improve neuronal survival

2010 ◽  
Vol 1353 ◽  
pp. 194-203 ◽  
Author(s):  
Jinny Jung Yoon ◽  
Louise Frances Basford Nicholson ◽  
Sheryl Xia Feng ◽  
Jose C. Vis ◽  
Colin Richard Green
Keyword(s):  
Brain Slice ◽  
Neuronal Survival ◽  
Antisense Oligodeoxynucleotides ◽  
Slice Culture ◽  
Novel Method ◽  
Organotypic Brain Slice ◽  
Brain Slice Culture

scholarly journals Organotypic Brain Slice Culture Microglia Exhibit Molecular Similarity to Acutely-Isolated Adult Microglia and Provide a Platform to Study Neuroinflammation

2020 ◽  
Vol 14 ◽  
Author(s):  
Alex R. D. Delbridge ◽  
Dann Huh ◽  
Margot Brickelmaier ◽  
Jeremy C. Burns ◽  
Chris Roberts ◽  
...  
Keyword(s):  
Gene Expression ◽  
Brain Slice ◽  
Initial Period ◽  
Slice Culture ◽  
Organotypic Brain Slice ◽  
Brain Slice Cultures ◽  
The Impact ◽  
Over Time

Microglia are central nervous system (CNS) resident immune cells that have been implicated in neuroinflammatory pathogenesis of a variety of neurological conditions. Their manifold context-dependent contributions to neuroinflammation are only beginning to be elucidated, which can be attributed in part to the challenges of studying microglia in vivo and the lack of tractable in vitro systems to study microglia function. Organotypic brain slice cultures offer a tissue-relevant context that enables the study of CNS resident cells and the analysis of brain slice microglial phenotypes has provided important insights, in particular into neuroprotective functions. Here we use RNA sequencing, direct digital quantification of gene expression with nCounter® technology and targeted analysis of individual microglial signature genes, to characterize brain slice microglia relative to acutely-isolated counterparts and 2-dimensional (2D) primary microglia cultures, a widely used in vitro surrogate. Analysis using single cell and population-based methods found brain slice microglia exhibited better preservation of canonical microglia markers and overall gene expression with stronger fidelity to acutely-isolated adult microglia, relative to in vitro cells. We characterized the dynamic phenotypic changes of brain slice microglia over time, after plating in culture. Mechanical damage associated with slice preparation prompted an initial period of inflammation, which resolved over time. Based on flow cytometry and gene expression profiling we identified the 2-week timepoint as optimal for investigation of microglia responses to exogenously-applied stimuli as exemplified by treatment-induced neuroinflammatory changes observed in microglia following LPS, TNF and GM-CSF addition to the culture medium. Altogether these findings indicate that brain slice cultures provide an experimental system superior to in vitro culture of microglia as a surrogate to investigate microglia functions, and the impact of soluble factors and cellular context on their physiology.

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