Journal articles: 'Organotypic Brain Slice Co-Cultures'

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Humpel, Christian. "Organotypic Brain Slice Cultures." Current Protocols in Immunology 123, no. 1 (October 12, 2018): e59. http://dx.doi.org/10.1002/cpim.59.

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Humpel, Christian. "Organotypic Brain Slices of ADULT Transgenic Mice: A Tool to Study Alzheimer’s Disease." Current Alzheimer Research 16, no. 2 (February 4, 2019): 172–81. http://dx.doi.org/10.2174/1567205016666181212153138.

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Transgenic mice have been extensively used to study the Alzheimer pathology. In order to reduce, refine and replace (3Rs) the number of animals, ex vivo cultures are used and optimized. Organotypic brain slices are the most potent ex vivo slice culture models, keeping the 3-dimensional structure of the brain and being closest to the in vivo situation. Organotypic brain slice cultures have been used for many decades but were mainly prepared from postnatal (day 8-10) old rats or mice. More recent work (including our lab) now aims to culture organotypic brain slices from adult mice including transgenic mice. Especially in Alzheimer´s disease research, brain slices from adult transgenic mice will be useful to study beta-amyloid plaques, tau pathology and glial activation. This review will summarize the studies using organotypic brain slice cultures from adult mice to mimic Alzheimer's disease and will highlight advantages and also pitfalls using this technique.

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Humpel, C. "Organotypic brain slice cultures: A review." Neuroscience 305 (October 2015): 86–98. http://dx.doi.org/10.1016/j.neuroscience.2015.07.086.

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Phillips, Wiktor S., Mikkel Herly, Christopher A. Del Negro, and Jens C. Rekling. "Organotypic slice cultures containing the preBötzinger complex generate respiratory-like rhythms." Journal of Neurophysiology 115, no. 2 (February 1, 2016): 1063–70. http://dx.doi.org/10.1152/jn.00904.2015.

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Study of acute brain stem slice preparations in vitro has advanced our understanding of the cellular and synaptic mechanisms of respiratory rhythm generation, but their inherent limitations preclude long-term manipulation and recording experiments. In the current study, we have developed an organotypic slice culture preparation containing the preBötzinger complex (preBötC), the core inspiratory rhythm generator of the ventrolateral brain stem. We measured bilateral synchronous network oscillations, using calcium-sensitive fluorescent dyes, in both ventrolateral (presumably the preBötC) and dorsomedial regions of slice cultures at 7–43 days in vitro. These calcium oscillations appear to be driven by periodic bursts of inspiratory neuronal activity, because whole cell recordings from ventrolateral neurons in culture revealed inspiratory-like drive potentials, and no oscillatory activity was detected from glial fibrillary associated protein-expressing astrocytes in cultures. Acute slices showed a burst frequency of 10.9 ± 4.2 bursts/min, which was not different from that of brain stem slice cultures (13.7 ± 10.6 bursts/min). However, slice cocultures that include two cerebellar explants placed along the dorsolateral border of the brainstem displayed up to 193% faster burst frequency (22.4 ± 8.3 bursts/min) and higher signal amplitude (340%) compared with acute slices. We conclude that preBötC-containing slice cultures retain inspiratory-like rhythmic function and therefore may facilitate lines of experimentation that involve extended incubation (e.g., genetic transfection or chronic drug exposure) while simultaneously being amenable to imaging and electrophysiology at cellular, synaptic, and network levels.

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Heine, Claudia, Katja Sygnecka, Nico Scherf, Marcus Grohmann, Annett Bräsigk, and Heike Franke. "P2Y1 receptor mediated neuronal fibre outgrowth in organotypic brain slice co-cultures." Neuropharmacology 93 (June 2015): 252–66. http://dx.doi.org/10.1016/j.neuropharm.2015.02.001.

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Joost, Sarah, Stefan Mikkat, Michael Wille, Antje Schümann, and Oliver Schmitt. "Membrane Protein Identification in Rodent Brain Tissue Samples and Acute Brain Slices." Cells 8, no. 5 (May 8, 2019): 423. http://dx.doi.org/10.3390/cells8050423.

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Acute brain slices are a sample format for electrophysiology, disease modeling, and organotypic cultures. Proteome analyses based on mass spectrometric measurements are seldom used on acute slices, although they offer high-content protein analyses and explorative approaches. In neuroscience, membrane proteins are of special interest for proteome-based analysis as they are necessary for metabolic, electrical, and signaling functions, including myelin maintenance and regeneration. A previously published protocol for the enrichment of plasma membrane proteins based on aqueous two-phase polymer systems followed by mass spectrometric protein identification was adjusted to the small sample size of single acute murine slices from newborn animals and the reproducibility of the results was analyzed. For this, plasma membrane proteins of 12 acute slice samples from six animals were enriched and analyzed by liquid chromatography-mass spectrometry. A total of 1161 proteins were identified, of which 369 were assigned to membranes. Protein abundances showed high reproducibility between samples. The plasma membrane protein separation protocol can be applied to single acute slices despite the low sample size and offers a high yield of identifiable proteins. This is not only the prerequisite for proteome analysis of organotypic slice cultures but also allows for the analysis of small-sized isolated brain regions at the proteome level.

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Ucar, Buket, Sedef Yusufogullari, and Christian Humpel. "Collagen hydrogels loaded with fibroblast growth factor-2 as a bridge to repair brain vessels in organotypic brain slices." Experimental Brain Research 238, no. 11 (August 29, 2020): 2521–29. http://dx.doi.org/10.1007/s00221-020-05907-7.

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Abstract Vessel damage is a general pathological process in many neurodegenerative disorders, as well as spinal cord injury, stroke, or trauma. Biomaterials can present novel tools to repair and regenerate damaged vessels. The aim of the present study is to test collagen hydrogels loaded with different angiogenic factors to study vessel repair in organotypic brain slice cultures. In the experimental set up I, we made a cut on the organotypic brain slice and tested re-growth of laminin + vessels. In the experimental set up II, we cultured two half brain slices with a gap with a collagen hydrogel placed in between to study endothelial cell migration. In the experimental set up I, we showed that the number of vessels crossing the cut was tendencially increased with the addition of fibroblast growth factor-2 (FGF-2), vascular endothelial growth factor, or platelet-derived growth factor-BB compared to the control group. In the experimental set up II, we demonstrated that a collagen hydrogel loaded with FGF-2 resulted in a significantly increased number of migrated laminin + cells in the gap between the slices compared to the control hydrogel. Co-administration of several growth factors did not further potentiate the effects. Taken together, we show that organotypic brain slices are good models to study brain vessels and FGF-2 is a potent angiogenic factor for endothelial cell proliferation and migration. Our results provide evidence that the collagen hydrogels can be used as an extracellular matrix for the vascular endothelial cells.

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GIANINAZZI, C., M. SCHILD, N. MÜLLER, S. L. LEIB, F. SIMON, S. NUÑEZ, P. JOSS, and B. GOTTSTEIN. "Organotypic slice cultures from rat brain tissue: a new approach forNaegleria fowleriCNS infectionin vitro." Parasitology 132, no. 6 (September 13, 2005): 797–804. http://dx.doi.org/10.1017/s0031182005008619.

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The free-living amoebaNaegleria fowleriis the aetiological agent of primary amoebic meningoencephalitis (PAM), a disease leading to death in the vast majority of cases. In patients suffering from PAM, and in corresponding animal models, the brain undergoes a massive inflammatory response, followed by haemorrhage and severe tissue necrosis. Both,in vivoandin vitromodels are currently being used to study PAM infection. However, animal models may pose ethical issues, are dependent upon availability of specific infrastructural facilities, and are time-consuming and costly. Conversely, cell cultures lack the complex organ-specific morphology foundin vivo, and thus, findings obtainedin vitrodo not necessarily reflect the situationin vivo. The present study reports infection of organotypic slice cultures from rat brain withN. fowleriand compares the findings in this culture system within vivoinfection in a rat model of PAM, that proved complementary to that of mice. We found that brain morphology, as presentin vivo, is well retained in organotypic slice cultures, and that infection time-course including tissue damage parallels the observationsin vivoin the rat. Therefore, organotypic slice cultures from rat brain offer a newin vitroapproach to studyN. fowleriinfection in the context of PAM.

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Spahr-Schopfer, Isabelle, Lazlo Vutskits, Nicholas Toni, Pierre-Alain Buchs, Lorena Parisi, and Dominique Muller. "Differential Neurotoxic Effects of Propofol on Dissociated Cortical Cells and Organotypic Hippocampal Cultures." Anesthesiology 92, no. 5 (May 1, 2000): 1408–17. http://dx.doi.org/10.1097/00000542-200005000-00032.

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Background Propofol is a widely used anesthetic agent for adults and children. Although extensive clinical use has demonstrated its safety, neurologic dysfunctions have been described after the use of this agent. A recent study on a model of aggregating cell cultures reported that propofol might cause irreversible lesions of gamma-aminobutyric acid-mediated (GABAergic) neurons when administered at a critical phase of brain development. We investigated this issue by comparing the effects of long-term propofol treatment on two models of brain cultures: dissociated neonatal cortical cell cultures and organotypic slice cultures. Methods Survival of GABAergic neurons in dissociated cultures of newborn rat cortex (postnatal age, 1 day) treated for 3 days with different concentrations of propofol was assessed using histologic and cytochemical methods. For hippocampal organotypic slice cultures (postnatal age, 1 and 7 days), cell survival was assessed by measuring functional and morphologic parameters: extracellular and intracellular electrophysiology, propidium staining of dying cells, and light and electron microscopy. Results In dissociated neonatal cell cultures, propofol induced dose-dependent lesions of GABAergic neurons and of glial cells. In contrast, no evidence for neurotoxic effects of propofol were found after long-term treatment of organotypic slice cultures. Excitatory transmission was not affected by propofol, and inhibitory transmission was still functional. Histologic preparations showed no evidence for cell degeneration or death. Conclusion Although long-term applications of propofol to dissociated cortical cell cultures produced degeneration and death of GABAergic neurons and glial cells, no such lesions were found when using a model of postnatal organotypic slice cultures. This conclusion is based on both functional and morphologic tests.

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Croft, Cara L., and Wendy Noble. "Preparation of organotypic brain slice cultures for the study of Alzheimer’s disease." F1000Research 7 (May 15, 2018): 592. http://dx.doi.org/10.12688/f1000research.14500.1.

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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.

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Croft, Cara L., and Wendy Noble. "Preparation of organotypic brain slice cultures for the study of Alzheimer’s disease." F1000Research 7 (June 27, 2018): 592. http://dx.doi.org/10.12688/f1000research.14500.2.

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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.

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Wilhelmi, Eckbert, Ulrich H. Schöder, Akilah Benabdallah, Frank Sieg, Jörg Breder, and Klaus G. Reymann. "Organotypic Brain-slice Cultures from Adult Rats: Approaches for a Prolonged Culture Time." Alternatives to Laboratory Animals 30, no. 3 (May 2002): 275–83. http://dx.doi.org/10.1177/026119290203000304.

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Animal experiments are widely used in neurobiological and neuropharmacological research. Today, juvenile brain organotypic slice cultures have partially replaced in vivo experiments, but there is no adequate in vitro counterpart for the adult brain. The present study was aimed at the long-term culture of physiologically intact hippocampal slices from adult rats, by improving the conditions for preparation and culture, and the development of a new culture medium. A cerebrospinal fluid (CSF)-like medium was used, which was modified with a variety of supplements, including energy precursors, free-radical scavengers, and compounds known to inhibit neurotoxicity. The population spike amplitude (PSA) was used as a measure of viability, and amplitudes larger than 1mV indicated viable cultures. The addition of MK-801 during slice preparation improved PSA values during the first two days in vitro (DIV). Ascorbic acid and insulin prolonged the culture time up to DIV 4. FK-506 and vitamin E, alone or in combination, supported slice culture up to DIV 5. An increase in ATP, unless combined with vitamin E, and/or insulin, increased culture time up to DIV 6. Vitamins B1, B2, B12 and D2 had no effect. The modified CSF-like medium developed in this study permits the culture of adult hippocampal tissue for at least 6 days.

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Sidorcenco, Vasile, Luisa Krahnen, Marion Schulz, Janina Remy, Donat Kögel, Achim Temme, Ute Krügel, Heike Franke, and Achim Aigner. "Glioblastoma Tissue Slice Tandem-Cultures for Quantitative Evaluation of Inhibitory Effects on Invasion and Growth." Cancers 12, no. 9 (September 21, 2020): 2707. http://dx.doi.org/10.3390/cancers12092707.

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Glioblastomas (GBMs) are the most malignant brain tumors and are essentially incurable even after extensive surgery, radiotherapy, and chemotherapy, mainly because of extensive infiltration of tumor cells into the adjacent normal tissue. Thus, the evaluation of novel drugs in malignant glioma treatment requires sophisticated ex vivo models that approach the authentic interplay between tumor and host environment while avoiding extensive in vivo studies in animals. This paper describes the standardized setup of an organotypic brain tissue slice tandem-culture system, comprising of normal brain tissue from adult mice and tumor tissue from human glioblastoma xenografts, and explore its utility for assessing inhibitory effects of test drugs. The microscopic analysis of vertical sections of the slice tandem-cultures allows for the simultaneous assessment of (i) the invasive potential of single cells or cell aggregates and (ii) the space occupying growth of the bulk tumor mass, both contributing to malignant tumor progression. The comparison of tissue slice co-cultures with spheroids vs. tissue slice tandem-cultures using tumor xenograft slices demonstrates advantages of the xenograft tandem approach. The direct and facile application of test drugs is shown to exert inhibitory effects on bulk tumor growth and/or tumor cell invasion, and allows their precise quantitation. In conclusion, we describe a straightforward ex vivo system mimicking the in vivo situation of the tumor mass and the normal brain in GBM patients. It reduces animal studies and allows for the direct and reproducible application of test drugs and the precise quantitation of their effects on the bulk tumor mass and on the tumor’s invasive properties.

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Morawski, Markus, Alexander Dityatev, Maike Hartlage-Rübsamen, Maren Blosa, Max Holzer, Katharina Flach, Sanja Pavlica, et al. "Tenascin-R promotes assembly of the extracellular matrix of perineuronal nets via clustering of aggrecan." Philosophical Transactions of the Royal Society B: Biological Sciences 369, no. 1654 (October 19, 2014): 20140046. http://dx.doi.org/10.1098/rstb.2014.0046.

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Perineuronal nets (PNs) in the brains of tenascin-R-deficient ( tn-r −/− ) mice develop in temporal concordance with those of wild-type ( tn-r +/+ ) mice. However, the histological appearance of PNs is abnormal in adult tn-r −/− mice. Here, we investigated whether similar defects are also seen in dissociated and organotypic cultures from hippocampus and forebrain of tn-r −/− mice and whether the structure of PNs could be normalized. In tn-r −/− cultures, accumulations of several extracellular matrix molecules were mostly associated with somata, whereas dendrites were sparsely covered, compared with tn-r +/+ mice. Experiments to normalize the structure of PNs in tn-r −/− organotypic slice cultures by depolarization of neurons, or by co-culturing tn-r +/+ and tn-r −/− brain slices failed to restore a normal PN phenotype. However, formation of dendritic PNs in cultures was improved by the application of tenascin-R protein and rescued by polyclonal antibodies to aggrecan and a bivalent, but not monovalent form of the lectin Wisteria floribunda agglutinin. These results show that tenascin-R and aggrecan are decisive contributors to formation and stabilization of PNs and that tenascin-R may implement these functions by clustering of aggrecan. Proposed approaches for restoration of normal PN structure are noteworthy in the context of PN abnormalities in neurological disorders, such as epilepsy, schizophrenia and addiction.

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Thomas, Mark P., Margaret I. Davis, Daniel T. Monaghan, and Richard A. Morrisett. "Organotypic Brain Slice Cultures for Functional Analysis of Alcohol-Related Disorders." Alcoholism: Clinical & Experimental Research 22, no. 1 (February 1998): 51. http://dx.doi.org/10.1097/00000374-199802000-00006.

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Falsig, Jeppe. "S5-01-06: Prion-induced neurodegeneration in organotypic brain slice cultures." Alzheimer's & Dementia 5, no. 4S_Part_6 (July 2009): P167. http://dx.doi.org/10.1016/j.jalz.2009.05.575.

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Noraberg, J., F. Poulsen, M. Blaabjerg, B. Kristensen, C. Bonde, M. Montero, M. Meyer, J. Gramsbergen, and J. Zimmer. "Organotypic Hippocampal Slice Cultures for Studies of Brain Damage, Neuroprotection and Neurorepair." Current Drug Target -CNS & Neurological Disorders 4, no. 4 (August 1, 2005): 435–52. http://dx.doi.org/10.2174/1568007054546108.

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Jahnsen, Henrik, Bjarne W. Kristensen, Pierre Thiébaud, Jens Noraberg, Birthe Jakobsen, Marco Bove, Sergio Martinoia, Milena Koudelka-Hep, Massimo Grattarola, and Jens Zimmer. "Coupling of Organotypic Brain Slice Cultures to Silicon-Based Arrays of Electrodes." Methods 18, no. 2 (June 1999): 160–72. http://dx.doi.org/10.1006/meth.1999.0769.

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Rappoldt, Liam, Adrienne Weeks, Rodney Ouellete, Jeremy Roy, Catherine Taylor, Craig McCormick, Kathleen Attwood, and Inhwa Kim. "TMOD-26. ESTABLISHING A PATIENT-DERIVED, IN-VITRO ORGANOTYPIC SLICE CULTURE MODEL OF GBM." Neuro-Oncology 22, Supplement_2 (November 2020): ii233. http://dx.doi.org/10.1093/neuonc/noaa215.976.

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Abstract Glioblastoma Multiforme (GBM) is the most common primary malignant brain tumour. This tumour is universally fatal with a median survival of 15 months. Driving this pathology is an extremely heterogeneic tumour and complex tumour microenvironment. GBM research is primarily conducted using immortalized or primary cell lines due to their practicality and reproducibility. However, these cell lines do not effectively recapitulate the tumour microenvironment. Mouse models address these shortcomings but are laborious and expensive. We propose to utilize a patient derived organotypic culture model of GBM as an intermediary. We have utilized this model to test genetic manipulation via lentiviral transduction and the feasibility of utilizing this model to understand patient derived extracellular vesicles (EVs). We have sectioned and cultured patient derived organotypic models for 14 days without loss of viability. To determine if these organotypic cultures are amenable to lentiviral manipulation, tissue sections were transduced with far-red fluorescent lentivirus and efficiency determined by confocal laser scanning microscopy (CLSM) and flow cytometry (FC). To determine feasibility as a model for EVs, media obtained from patient-derived organotypic cultures was analyzed by western blot, nanoparticle tracking analysis (NTA), and nanoFlow Cytometry (nFC). In the future these EVs will be compared to those found in patient serum. The model of GBM has been lentivirally transduced to express a far-red fluorescent vector in approximately 15% of the slice, quantified by CLSM and FC. EV-sized particles positive for canonical EV markers have been identified in the media by NTA, nFC and western blot. Using lentiviral-mediated genetic engineering and emerging EV science, this organotypic slice culture models yields exciting utility in GBM research. The established organotypic slice culture model will likely be a valuable tool in the study of GBM biology and EV dynamics.

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Laksono, Brigitta M., Diana N. Tran, Ivanela Kondova, Harry G. H. van Engelen, Samira Michels, Sham Nambulli, Rory D. de Vries, W. Paul Duprex, Georges M. G. M. Verjans, and Rik L. de Swart. "Comparable Infection Level and Tropism of Measles Virus and Canine Distemper Virus in Organotypic Brain Slice Cultures Obtained from Natural Host Species." Viruses 13, no. 8 (August 10, 2021): 1582. http://dx.doi.org/10.3390/v13081582.

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Measles virus (MV) and canine distemper virus (CDV) are closely related members of the family Paramyxoviridae, genus Morbillivirus. MV infection of humans and non-human primates (NHPs) results in a self-limiting disease, which rarely involves central nervous system (CNS) complications. In contrast, infection of carnivores with CDV usually results in severe disease, in which CNS complications are common and the case-fatality rate is high. To compare the neurovirulence and neurotropism of MV and CDV, we established a short-term organotypic brain slice culture system of the olfactory bulb, hippocampus, or cortex obtained from NHPs, dogs, and ferrets. Slices were inoculated ex vivo with wild-type-based recombinant CDV or MV expressing a fluorescent reporter protein. The infection level of both morbilliviruses was determined at different times post-infection. We observed equivalent infection levels and identified microglia as main target cells in CDV-inoculated carnivore and MV-inoculated NHP brain tissue slices. Neurons were also susceptible to MV infection in NHP brain slice cultures. Our findings suggest that MV and CDV have comparable neurotropism and intrinsic capacity to infect CNS-resident cells of their natural host species.

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Sanders, Robert D., Jing Xu, Yi Shu, Adam Januszewski, Sunil Halder, Antonio Fidalgo, Pamela Sun, Mahmuda Hossain, Daqing Ma, and Mervyn Maze. "Dexmedetomidine Attenuates Isoflurane-induced Neurocognitive Impairment in Neonatal Rats." Anesthesiology 110, no. 5 (May 1, 2009): 1077–85. http://dx.doi.org/10.1097/aln.0b013e31819daedd.

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Background Neuroapoptosis is induced by the administration of anesthetic agents to the young. As alpha2 adrenoceptor signaling plays a trophic role during development and is neuroprotective in several settings of neuronal injury, the authors investigated whether dexmedetomidine could provide functional protection against isoflurane-induced injury. Methods Isoflurane-induced injury was provoked in organotypic hippocampal slice cultures in vitro or in vivo in postnatal day 7 rats by a 6-h exposure to 0.75% isoflurane with or without dexmedetomidine. In vivo, the alpha2 adrenoceptor antagonist atipamezole was used to identify if dexmedetomidine neuroprotection involved alpha2 adrenoceptor activation. The gamma-amino-butyric-acid type A antagonist, gabazine, was also added to the organotypic hippocampal slice cultures in the presence of isoflurane. Apoptosis was assessed using cleaved caspase-3 immunohistochemistry. Cognitive function was assessed in vivo on postnatal day 40 using fear conditioning. Results In vivo dexmedetomidine dose-dependently prevented isoflurane-induced injury in the hippocampus, thalamus, and cortex; this neuroprotection was attenuated by treatment with atipamezole. Although anesthetic treatment did not affect the acquisition of short-term memory, isoflurane did induce long-term memory impairment. This neurocognitive deficit was prevented by administration of dexmedetomidine, which also inhibited isoflurane-induced caspase-3 expression in organotypic hippocampal slice cultures in vitro; however, gabazine did not modify this neuroapoptosis. Conclusion Dexmedetomidine attenuates isoflurane-induced injury in the developing brain, providing neurocognitive protection. Isoflurane-induced injury in vitro appears to be independent of activation of the gamma-amino-butyric-acid type A receptor. If isoflurane-induced neuroapoptosis proves to be a clinical problem, administration of dexmedetomidine may be an important adjunct to prevent isoflurane-induced neurotoxicity.

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Duport, S., F. Robert, D. Muller, G. Grau, L. Parisi, and L. Stoppini. "An in vitro blood-brain barrier model: Cocultures between endothelial cells and organotypic brain slice cultures." Proceedings of the National Academy of Sciences 95, no. 4 (February 17, 1998): 1840–45. http://dx.doi.org/10.1073/pnas.95.4.1840.

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Kristensen, Bjarne W., Jens Noraberg, Pierre Thiébaud, Milena Koudelka-Hep, and Jens Zimmer. "Biocompatibility of silicon-based arrays of electrodes coupled to organotypic hippocampal brain slice cultures." Brain Research 896, no. 1-2 (March 2001): 1–17. http://dx.doi.org/10.1016/s0006-8993(00)03304-7.

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McCaughey-Chapman, Amy, and Bronwen Connor. "Rat brain sagittal organotypic slice cultures as an ex vivo dopamine cell loss system." Journal of Neuroscience Methods 277 (February 2017): 83–87. http://dx.doi.org/10.1016/j.jneumeth.2016.12.012.

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Xu, Bin, Sheng-Wen Wu, Chun-Wei Lu, Yu Deng, Wei Liu, Yan-Gang Wei, Tian-Yao Yang, and Zhao-Fa Xu. "Oxidative stress involvement in manganese-induced alpha-synuclein oligomerization in organotypic brain slice cultures." Toxicology 305 (March 2013): 71–78. http://dx.doi.org/10.1016/j.tox.2013.01.006.

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Guldimann, Claudia, Beatrice Lejeune, Sandra Hofer, Stephen L. Leib, Joachim Frey, Andreas Zurbriggen, Torsten Seuberlich, and Anna Oevermann. "Ruminant organotypic brain-slice cultures as a model for the investigation of CNS listeriosis." International Journal of Experimental Pathology 93, no. 4 (July 18, 2012): 259–68. http://dx.doi.org/10.1111/j.1365-2613.2012.00821.x.

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Mewes, Agneta, Heike Franke, and David Singer. "Organotypic Brain Slice Cultures of Adult Transgenic P301S Mice—A Model for Tauopathy Studies." PLoS ONE 7, no. 9 (September 11, 2012): e45017. http://dx.doi.org/10.1371/journal.pone.0045017.

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Guldimann, C., A. Zurbriggen, B. Lejeune, T. Seuberlich, and A. Oevermann. "Organotypic Brain Slice Cultures as a Tool for the Investigation of Listeriosis in Ruminants." Journal of Comparative Pathology 146, no. 1 (January 2012): 60. http://dx.doi.org/10.1016/j.jcpa.2011.11.061.

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Xiao, Li, Chikako Saiki, and Hisashi Okamura. "Oxidative Stress-Tolerant Stem Cells from Human Exfoliated Deciduous Teeth Decrease Hydrogen Peroxide-Induced Damage in Organotypic Brain Slice Cultures from Adult Mice." International Journal of Molecular Sciences 20, no. 8 (April 15, 2019): 1858. http://dx.doi.org/10.3390/ijms20081858.

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Oxidative stress causes severe tissue injury of the central nervous system in ischemic brain damage (IBD), traumatic brain injury (TBI) and neurodegenerative disorders. In this study, we used hydrogen peroxide (H2O2) to induce oxidative stress in organotypic brain slice cultures (OBSCs), and investigated the protective effects of oxidative stress-tolerant (OST) stem cells harvested from human exfoliated deciduous teeth (SHED) which were co-cultivated with OBSCs. Using presto blue assay and immunostaining, we demonstrated that both normal SHED and OST-SHED could prevent H2O2-induced cell death, and increase the numbers of mature neuron and neuronal progenitors in the hippocampus of OBSCs. During co-cultivation, OST-SHED, but not normal SHED, exhibited neuronal cell morphology and expressed neuronal markers. Results from ELISA showed that both normal SHED and OST-SHED significantly decreased oxidative DNA damage in H2O2-treated OBSCs. SHED could also produce neurotrophic factor BDNF (brain derived neurotrophic factor) and promoted the production of IL-6 in OBSCs. Although OST-SHED had lower cell viability, the neuronal protection of OST-SHED was significantly superior to that of normal SHED. Our findings suggest that SHED, especially OST-SHED, could prevent oxidative stress induced brain damage. OST-SHED can be explored as a new therapeutic tool for IBD, TBI and neurodegenerative disorders.

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Lange-Asschenfeldt, Christian, Ami P. Raval, Kunjan R. Dave, Daria Mochly-Rosen, Thomas J. Sick, and Miguel A. Pérez-Pinzón. "Epsilon Protein Kinase C Mediated Ischemic Tolerance Requires Activation of the Extracellular Regulated Kinase Pathway in the Organotypic Hippocampal Slice." Journal of Cerebral Blood Flow & Metabolism 24, no. 6 (June 2004): 636–45. http://dx.doi.org/10.1097/01.wcb.0000121235.42748.bf.

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Ischemic preconditioning (IPC) promotes brain tolerance against subsequent ischemic insults. Using the organotypic hippocampal slice culture, we conducted the present study to investigate (1) the role of adenosine A1 receptor (A1AR) activation in IPC induction, (2) whether epsilon protein kinase C (ɛPKC) activation after IPC is mediated by the phosphoinositol pathway, and (3) whether ɛPKC protection is mediated by the extracellular signal-regulated kinase (ERK) pathway. Our results demonstrate that activation of A1AR emulated IPC, whereas blockade of the A1AR during IPC diminished neuroprotection. The neuroprotection promoted by the A1AR was also reduced by the ɛPKC antagonist. To determine whether ɛPKC activation in IPC and A1AR preconditioning is mediated by activation of the phosphoinositol pathway, we incubated slices undergoing IPC or adenosine treatment with a phosphoinositol phospholipase C inhibitor. In both cases, preconditioning neuroprotection was significantly attenuated. To further characterize the subsequent signal transduction pathway that ensues after ɛPKC activation, mitogen-activated protein kinase kinase was blocked during IPC and pharmacologic preconditioning (PPC) (with ɛPKC, NMDA, or A1AR agonists). This treatment significantly attenuated IPC- and PPC-induced neuroprotection. In conclusion, we demonstrate that ɛPKC activation after IPC/PPC is essential for neuroprotection against oxygen/glucose deprivation in organotypic slice cultures and that the ERK pathway is downstream to ɛPKC.

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Marques-Torrejon, Maria Angeles, Ester Gangoso, and Steven M. Pollard. "Modelling glioblastoma tumour-host cell interactions using adult brain organotypic slice co-culture." Disease Models & Mechanisms 11, no. 2 (December 1, 2017): dmm031435. http://dx.doi.org/10.1242/dmm.031435.

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Renic, Marija, Suresh N. Kumar, Debebe Gebremedhin, Matthew A. Florence, Nashaat Z. Gerges, John R. Falck, David R. Harder, and Richard J. Roman. "Protective effect of 20-HETE inhibition in a model of oxygen-glucose deprivation in hippocampal slice cultures." American Journal of Physiology-Heart and Circulatory Physiology 302, no. 6 (March 15, 2012): H1285—H1293. http://dx.doi.org/10.1152/ajpheart.00340.2011.

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Recent studies have indicated that inhibitors of the synthesis of 20-hydroxyeicosatetraenoic acid (20-HETE) may have direct neuroprotective actions since they reduce infarct volume after ischemia reperfusion in the brain without altering blood flow. To explore this possibility, the present study used organotypic hippocampal slice cultures subjected to oxygen-glucose deprivation (OGD) and reoxygenation to examine whether 20-HETE is released by organotypic hippocampal slices after OGD and whether it contributes to neuronal death through the generation of ROS and activation of caspase-3. The production of 20-HETE increased twofold after OGD and reoxygenation. Blockade of the synthesis of 20-HETE with N-hydroxy- N′-(4-butyl-2-methylphenol)formamidine (HET0016) or its actions with a 20-HETE antagonist, 20-hydroxyeicosa-6( Z),15( Z)-dienoic acid, reduced cell death, as measured by the release of lactate dehydrogenase and propidium iodide uptake. Administration of a 20-HETE mimetic, 20-hydroxyeicosa-5( Z),14( Z)-dienoic acid (5,14-20-HEDE), had the opposite effect and increased injury after OGD. The death of neurons after OGD was associated with an increase in the production of ROS and activation of caspase-3. These effects were attenuated by HET0016 and potentiated after the administration of 5,14-20-HEDE. These findings indicate that the production of 20-HETE by hippocampal slices is increased after OGD and that inhibitors of the synthesis or actions of 20-HETE protect neurons from ischemic cell death. The protective effect of 20-HETE inhibitors is associated with a decrease in superoxide production and activation of caspase-3.

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Zehendner, Christoph M., Heiko J. Luhmann, and Christoph RW Kuhlmann. "Studying the Neurovascular Unit: An Improved Blood–Brain Barrier Model." Journal of Cerebral Blood Flow & Metabolism 29, no. 12 (July 29, 2009): 1879–84. http://dx.doi.org/10.1038/jcbfm.2009.103.

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The blood–brain barrier (BBB) closely interacts with the neuronal parenchyma in vivo. To replicate this interdependence in vitro, we established a murine coculture model composed of brain endothelial cell (BEC) monolayers with cortical organotypic slice cultures. The morphology of cell types, expression of tight junctions, formation of reactive oxygen species, caspase-3 activity in BECs, and alterations of electrical resistance under physiologic and pathophysiological conditions were investigated. This new BBB model allows the application of techniques such as laser scanning confocal microscopy, immunohistochemistry, fluorescent live cell imaging, and electrical cell substrate impedance sensing in real time for studying the dynamics of BBB function under defined conditions.

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Hiolski, E. M., S. Ito, J. M. Beggs, K. A. Lefebvre, A. M. Litke, and D. R. Smith. "Domoic acid disrupts the activity and connectivity of neuronal networks in organotypic brain slice cultures." NeuroToxicology 56 (September 2016): 215–24. http://dx.doi.org/10.1016/j.neuro.2016.08.004.

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Thomas, Mark P., Margaret I. Davis, Daniel T. Monaghan, and Richard A. Morrisett. "Organotypic Brain Slice Cultures for Functional Analysis of Alcohol-Related Disorders: Novel Versus Conventional Preparations." Alcoholism: Clinical and Experimental Research 22, no. 1 (February 1998): 51–59. http://dx.doi.org/10.1111/j.1530-0277.1998.tb03616.x.

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Croft, Cara L., Matthew A. Wade, Ksenia Kurbatskaya, Pavlina Mastrandreas, Martina M. Hughes, Emma C. Phillips, Amy M. Pooler, Michael S. Perkinton, Diane P. Hanger, and Wendy Noble. "Membrane association and release of wild-type and pathological tau from organotypic brain slice cultures." Cell Death & Disease 8, no. 3 (March 2017): e2671-e2671. http://dx.doi.org/10.1038/cddis.2017.97.

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Noraberg, Jens. "Organotypic Brain Slice Cultures: An Efficient and Reliable Method for Neurotoxicological Screening and Mechanistic Studies." Alternatives to Laboratory Animals 32, no. 4 (October 2004): 329–37. http://dx.doi.org/10.1177/026119290403200403.

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Zimmer, J., B. W. Kristensen, B. Jakobsen, and J. Noraberg. "Excitatory amino acid neurotoxicity and modulation of glutamate receptor expression in organotypic brain slice cultures." Amino Acids 19, no. 1 (August 31, 2000): 7–21. http://dx.doi.org/10.1007/s007260070029.

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Förster, Eckart, Shanting Zhao, and Michael Frotscher. "Hyaluronan-associated adhesive cues control fiber segregation in the hippocampus." Development 128, no. 15 (August 1, 2001): 3029–39. http://dx.doi.org/10.1242/dev.128.15.3029.

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In various brain regions, particularly in the hippocampus, afferent fiber projections terminate in specific layers. Little is known about the molecular cues governing this laminar specificity. To this end we have recently shown that the innervation pattern of entorhinal fibers to the hippocampus is mimicked by the lamina-specific adhesion of entorhinal cells on living hippocampal slices, suggesting a role of adhesion molecules in the positioning of entorhinal fibers. Here, we have analyzed the role of extracellular matrix components in mediating this lamina-specific adhesion. We show that hyaluronidase treatment of hippocampal slices abolishes lamina-specific adhesion as well as layer-specific growth of entorhinal fibers to the dentate outer molecular layer in organotypic slice cultures. We conclude that hyaluronan-associated molecules play a crucial role in the formation of the lamina-specific entorhinal projection to the hippocampus.

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Rakotomamonjy, Jennifer, and Abdel Ghoumari. "Brain-Derived Neurotrophic Factor Is Required for the Neuroprotective Effect of Mifepristone on Immature Purkinje Cells in Cerebellar Slice Culture." International Journal of Molecular Sciences 20, no. 2 (January 12, 2019): 285. http://dx.doi.org/10.3390/ijms20020285.

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Endogenous γ-aminobutyric acid (GABA)-dependent activity induces death of developing Purkinje neurons in mouse organotypic cerebellar cultures and the synthetic steroid mifepristone blocks this effect. Here, using brain-derived neurotrophic factor (BDNF) heterozygous mice, we show that BDNF plays no role in immature Purkinje cell death. However, interestingly, BDNF haploinsufficiency impairs neuronal survival induced by mifepristone and GABAA-receptors antagonist (bicuculline) treatments, indicating that the underlying neuroprotective mechanism requires the neurotrophin full expression.

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Morrison, Barclay, David F. Meaney, Susan S. Margulies, and Tracy K. McIntosh. "Dynamic Mechanical Stretch of Organotypic Brain Slice Cultures Induces Differential Genomic Expression: Relationship to Mechanical Parameters." Journal of Biomechanical Engineering 122, no. 3 (February 6, 2000): 224–30. http://dx.doi.org/10.1115/1.429650.

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Although the material properties of biological tissues are reasonably well established, recent studies have suggested that the biological response of brain tissue and its constituent cells may also be viscoelastic and sensitive to both the magnitude and rate of a mechanical stimulus. Given the potential involvement of changes in gene expression in the pathogenic sequelae after head trauma, we analyzed the expression of 22 genes related to cell death and survival and found that a number of these genes were differentially regulated after mechanical stretch of an organotypic brain slice culture. Twenty-four hours after stretch, the expression of BDNF, NGF, and TrkA was significantly increased, whereas that of bcl-2, CREB, and GAD65 was significantly decreased (MANOVA followed by ANOVA, p<0.05). Expression of CREB and GAD65 was negatively correlated with strain, whereas expression of APP695 was negatively correlated with strain rate (all p<0.05). This study demonstrates that a subset of genes involved in cell death and survival are differentially regulated after dynamic stretch in vitro and that the expression of specific genes is correlated with mechanical parameters of that stretch. [S0148-0731(00)00303-4]

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Moelgg, Kurt, Faryal Jummun, and Christian Humpel. "Spreading of Beta-Amyloid in Organotypic Mouse Brain Slices and Microglial Elimination and Effects on Cholinergic Neurons." Biomolecules 11, no. 3 (March 15, 2021): 434. http://dx.doi.org/10.3390/biom11030434.

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The extracellular deposition of β-amyloid (Aβ) is one of the major characteristics in Alzheimer´s disease (AD). The “spreading hypothesis” suggests that a pathological protein (similar to prions) spreads over the entire brain. The aim of the present study was to use organotypic brain slices of postnatal day 8–10 mice. Using collagen hydrogels, we applied different Aβ peptides onto brain slices and analyzed spreading as well as glial reactions after eight weeks of incubation. Our data showed that from all tested Aβ peptides, human Aβ42 had the most potent activity to spread over into adjacent “target” areas. This effect was potentiated when brain slices from transgenic AD mice (APP_SweDI) were cultured. When different brain areas were connected to the “target slice” the spreading activity was more intense, originating from ventral striatum and brain stem. Reactive glial-fibrillary acidic protein (GFAP) astrogliosis increased over time, but Aβ depositions co-localized only with Iba1+ microglia but not with astrocytes. Application of human Aβ42 did not cause a degeneration of cholinergic neurons. We concluded that human Aβ42 spreads over into other “target areas”, causing activation of glial cells. Most of the spread Aβ42 was taken up by microglia, and thus toxic free Aβ could not damage cholinergic neurons.

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Kovács, Richard, Ismini Papageorgiou, and Uwe Heinemann. "Slice Cultures as a Model to Study Neurovascular Coupling and Blood Brain Barrier In Vitro." Cardiovascular Psychiatry and Neurology 2011 (February 16, 2011): 1–9. http://dx.doi.org/10.1155/2011/646958.

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Proper neuronal functioning depends on a strictly regulated interstitial environment and tight coupling of neuronal and metabolic activity involving adequate vascular responses. These functions take place at the blood brain barrier (BBB) composed of endothelial cells, basal lamina covered with pericytes, and the endfeet of perivascular astrocytes. In conventional in vitro models of the BBB, some of these components are missing. Here we describe a new model system for studying BBB and neurovascular coupling by using confocal microscopy and fluorescence staining protocols in organotypic hippocampal slice cultures. An elaborated network of vessels is retained in culture in spite of the absence of blood flow. Application of calcein-AM either from the interstitial or from the luminal side resulted in different staining patterns indicating the maintenance of a barrier. By contrast, the ethidium derivative MitoSox penetrated perivascular basal lamina and revealed free radical formation in contractile cells embracing the vessels, likely pericytes.

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Quadros, Amita, Nikunj Patel, Robert Crescentini, Fiona Crawford, Daniel Paris, and Michael Mullan. "Increased TNFα production and Cox-2 activity in organotypic brain slice cultures from APPsw transgenic mice." Neuroscience Letters 353, no. 1 (December 2003): 66–68. http://dx.doi.org/10.1016/j.neulet.2003.08.076.

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Xu, Bin, Ming Shan, Fei Wang, Yu Deng, Wei Liu, Shu Feng, Tian-Yao Yang, and Zhao-Fa Xu. "Endoplasmic reticulum stress signaling involvement in manganese-induced nerve cell damage in organotypic brain slice cultures." Toxicology Letters 222, no. 3 (October 2013): 239–46. http://dx.doi.org/10.1016/j.toxlet.2013.08.001.

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Reimers, Sabrina, Maike Hartlage-Rübsamen, Gert Brückner, and Steffen Roßner. "Formation of perineuronal nets in organotypic mouse brain slice cultures is independent of neuronal glutamatergic activity." European Journal of Neuroscience 25, no. 9 (June 6, 2007): 2640–48. http://dx.doi.org/10.1111/j.1460-9568.2007.05514.x.

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Gerace, Elisabetta, Elisa Landucci, Arianna Totti, Daniele Bani, Daniele Guasti, Roberto Baronti, Flavio Moroni, Guido Mannaioni, and Domenico E. Pellegrini-Giampietro. "Ethanol Toxicity During Brain Development: Alterations of Excitatory Synaptic Transmission in Immature Organotypic Hippocampal Slice Cultures." Alcoholism: Clinical and Experimental Research 40, no. 4 (April 2016): 706–16. http://dx.doi.org/10.1111/acer.13006.

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Wu, Qihui, Muhammad A. Shaikh, Emily S. Meymand, Bin Zhang, Kelvin C. Luk, John Q. Trojanowski, and Virginia M. Y. Lee. "Neuronal activity modulates alpha-synuclein aggregation and spreading in organotypic brain slice cultures and in vivo." Acta Neuropathologica 140, no. 6 (October 6, 2020): 831–49. http://dx.doi.org/10.1007/s00401-020-02227-6.

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Suckert, Theresa, Treewut Rassamegevanon, Johannes Müller, Antje Dietrich, Antonia Graja, Michael Reiche, Steffen Löck, Mechthild Krause, Elke Beyreuther, and Cläre von Neubeck. "Applying Tissue Slice Culture in Cancer Research—Insights from Preclinical Proton Radiotherapy." Cancers 12, no. 6 (June 16, 2020): 1589. http://dx.doi.org/10.3390/cancers12061589.

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A challenge in cancer research is the definition of reproducible, reliable, and practical models, which reflect the effects of complex treatment modalities and the heterogeneous response of patients. Proton beam radiotherapy (PBRT), relative to conventional photon-based radiotherapy, offers the potential for iso-effective tumor control, while protecting the normal tissue surrounding the tumor. However, the effects of PBRT on the tumor microenvironment and the interplay with newly developed chemo- and immunotherapeutic approaches are still open for investigation. This work evaluated thin-cut tumor slice cultures (TSC) of head and neck cancer and organotypic brain slice cultures (OBSC) of adult mice brain, regarding their relevance for translational radiooncology research. TSC and OBSC were treated with PBRT and investigated for cell survival with a lactate dehydrogenase (LDH) assay, DNA repair via the DNA double strand break marker γH2AX, as well as histology with regards to morphology. Adult OBSC failed to be an appropriate model for radiobiological research questions. However, histological analysis of TSC showed DNA damage and tumor morphological results, comparable to known in vivo and in vitro data, making them a promising model to study novel treatment approaches in patient-derived xenografts or primary tumor material.

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Kasparov, S., A. G. Teschemacher, and J. F. R. Paton. "Dynamic Confocal Imaging in Acute Brain Slices and Organotypic Slice Cultures Using a Spectral Confocal Microscope with Single Photon Excitation." Experimental Physiology 87, no. 6 (November 2002): 715–24. http://dx.doi.org/10.1113/eph8702480.

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Journal articles on the topic 'Organotypic Brain Slice Co-Cultures'

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