Mammalia

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    Protocols in Current Issue
    Isolation of Neural Stem Cells from the Embryonic Mouse Hippocampus for in vitro Growth or Engraftment into a Host Tissue
    Authors:  Oksana Rybachuk, Olga Kopach, Tetyana Pivneva and Vitaliy Kyryk, date: 02/20/2019, view: 713, Q&A: 0
    [Abstract] For both stem cell research and treatment of the central nervous system disorders, neural stem/progenitor cells (NSPCs) represent an important breakthrough tool. In the expanded stem cell-based therapy use, NSPCs not only provide a powerful cell source for neural cell replacement but a useful model for developmental biology research. Despite ...
    Collagenase-based Single Cell Isolation of Primary Murine Brain Endothelial Cells Using Flow Cytometry
    Authors:  Cathrin J. Czupalla, Hanadie Yousef, Tony Wyss-Coray and Eugene C. Butcher, date: 11/20/2018, view: 2057, Q&A: 0
    [Abstract] The brain endothelium is a highly specialized vascular structure that maintains the activity and integrity of the central nervous system (CNS). Previous studies have reported that the integrity of the brain endothelium is compromised in a plethora of neuropathologies. Therefore, it is of particular interest to establish a method that enables ...
    Papain-based Single Cell Isolation of Primary Murine Brain Endothelial Cells Using Flow Cytometry
    Authors:  Hanadie Yousef, Cathrin J Czupalla, Davis Lee, Eugene C Butcher and Tony Wyss-Coray, date: 11/20/2018, view: 1761, Q&A: 0
    [Abstract] Brain endothelial cells (BECs) form the integral component of the blood-brain barrier (BBB) which separates the systemic milieu from the brain parenchyma and protects the brain from pathogens and circulating factors. In order to study BEC biology, it was of particular interest to establish a method that enables researchers to investigate and ...
    Phagocytosis Assay for α-Synuclein Fibril Uptake by Mouse Primary Microglia
    Authors:  Cuilian Du, Feifan Zhang and Claire Xi Zhang, date: 09/05/2018, view: 1155, Q&A: 0
    [Abstract] Microglia are professional phagocytes in the brain and deficiency in their phagocytic activity plays an important role in Parkinson’s disease. This protocol mainly describes the phagocytosis assay for uptake of α-synuclein preformed fibrils, a pathologic form of α-synuclein, by primary microglia.
    Investigating Neural Stem Cell and Glioma Stem Cell Self-renewal Potential Using Extreme Limiting Dilution Analysis (ELDA)
    Authors:  Hong PT Nguyen, Paul M Daniel, Gulay Filiz and Theo Mantamadiotis, date: 09/05/2018, view: 1468, Q&A: 0
    [Abstract] Glioma stem cells (GSC) grown as neurospheres exhibit similar characteristics to neural stem cells (NSC) grown as neurospheres, including the ability to self-renew and differentiate. GSCs are thought to play a role in cancer initiation and progression. Self-renewal potential of GSCs is thought to reflect many characteristics associated with ...
    Preparation of Cerebellum Granule Neurons from Mouse or Rat Pups and Evaluation of Clostridial Neurotoxin Activity and Their Inhibitors by Western Blot and Immunohistochemistry
    Authors:  Domenico Azarnia Tehran and Marco Pirazzini, date: 07/05/2018, view: 2449, Q&A: 0
    [Abstract] Cerebellar Granule Neurons (CGN) from post-natal rodents have been widely used as a model to study neuronal development, physiology and pathology. CGN cultured in vitro maintain the same features displayed in vivo by mature cerebellar granule cells, including the development of a dense neuritic network, neuronal activity, ...
    Intracellular and Mitochondrial Reactive Oxygen Species Measurement in Primary Cultured Neurons
    [Abstract] Reactive oxygen species (ROS) are chemically reactive oxygen containing molecules. ROS consist of radical oxygen species including superoxide anion (O2•−) and hydroxyl radical (•OH) and non-radical oxygen species such as hydrogen peroxide (H2O2), singlet oxygen (O2). ROS are generated by ...
    Culture and Nucleofection of Postnatal Day 7 Cortical and Cerebellar Mouse Astroglial Cells
    Authors:  Malek Chouchane and Marcos Romualdo Costa, date: 02/05/2018, view: 2403, Q&A: 0
    [Abstract] Lineage reprogramming of astroglial cells isolated from different brain regions leads to the generation of different neuronal subtypes. This protocol describes the isolation and culture of neocortical and cerebellar astrocytes from postnatal mice. We also present a comprehensive description of the main steps towards successful gene delivery in ...
    Organotypic Brain Cultures: A Framework for Studying CNS Infection by Neurotropic Viruses and Screening Antiviral Drugs
    [Abstract] According to the World Health Organization (WHO), at least 50% of emerging viruses endowed with pathogenicity in humans can infect the Central Nervous System (CNS) with induction of encephalitis and other neurologic diseases (Taylor et al., 2001; Olival and Daszak, 2005). While neurological diseases are progressively documented, the ...
    Isolation, Culture and Differentiation of Adult Hippocampal Precursor Cells
    Authors:  Stefanie N. Bernas, Odette Leiter, Tara L. Walker and Gerd Kempermann, date: 11/05/2017, view: 5487, Q&A: 0
    [Abstract] There are two neurogenic niches in the adult mammalian brain: the subventricular zone of the lateral ventricle and the subgranular zone of the hippocampal dentate gyrus. Cells from these areas can be isolated and maintained in vitro, using two different culture systems to assess their potential regarding proliferation and differentiation ...



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