Cell Biology

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    Protocols in Current Issue
    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: 228, 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: 239, 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 ...
    HIVGKO: A Tool to Assess HIV-1 Latency Reversal Agents in Human Primary CD4+ T Cells
    Authors:  Emilie Battivelli and Eric Verdin, date: 10/20/2018, view: 324, Q&A: 0
    [Abstract] While able to suppress HIV replication in HIV infected individuals, combination antiretroviral therapy (ART) fails to eliminate viral latent reservoir, which consists in integrated transcriptional silenced HIV provirus. So far, identification of latently-infected cells has relied on activating cells to induce expression of HIV proteins which can ...
    Isolation and Culture of Mouse Lung ILC2s
    Authors:  Pauline Schmitt, Anais Duval, Emilie Mirey, Jean-Philippe Girard and Corinne Cayrol, date: 10/05/2018, view: 471, Q&A: 0
    [Abstract] Group 2 Innate Lymphoid Cells (ILC2) play an important role in immune responses at barrier surfaces, notably in the lung during airway allergic inflammation or asthma. Several studies have described methods to isolate ILC2s from wild-type naive mice, most of them using cell sorting to obtain a pure population. Here, we describe in detail, a ...
    Platelet Migration and Bacterial Trapping Assay under Flow
    Authors:  Shuxia Fan, Michael Lorenz, Steffen Massberg and Florian Gaertner, date: 09/20/2018, view: 529, Q&A: 0
    [Abstract] Blood platelets are critical for hemostasis and thrombosis, but also play diverse roles during immune responses. We have recently reported that platelets migrate at sites of infection in vitro and in vivo. Importantly, platelets use their ability to migrate to collect and bundle fibrin (ogen)-bound bacteria accomplishing ...
    Isolation of LYVE-1+ Endothelial Cells from Mouse Embryos
    Author:  Patrick Crosswhite, date: 08/05/2018, view: 488, Q&A: 0
    [Abstract] Lymphatic vessel endothelial hyaluronan receptor 1, or LYVE-1, is a type 1 integral membrane glycoprotein expressed by lymphatic endothelial cells (LECs). LYVE-1 is commonly used as a biological marker to visually distinguish developing lymphatic vessels from blood endothelial cells (arteries or veins). As our understanding of lymphatic biology is ...
    Measuring Secretion of Capsidiol in Leaf Tissues of Nicotiana benthamiana
    Authors:  Teruhiko Kuroyanagi, Maurizio Camagna and Daigo Takemoto, date: 08/05/2018, view: 560, Q&A: 0
    [Abstract] Plant species produce a wide variety of antimicrobial metabolites to protect themselves against potential pathogens in natural environments. Phytoalexins are low molecular weight compounds produced by plants in response to attempted attacks of pathogens. Accumulation of phytoalexins in attacked plant tissues can inhibit the growth of penetrating ...
    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: 1121, 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, ...
    Isolation of Microvascular Endothelial Cells
    Authors:  Kenneth C.P. Cheung and Federica M. Marelli-Berg, date: 06/20/2018, view: 1644, Q&A: 0
    [Abstract] The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. Murine endothelial cell culture is an important tool for cardiovascular disease research. This protocol demonstrates a quick, efficient method for the isolation of microvascular endothelial cells from murine ...
    Alphavirus Purification Using Low-speed Spin Centrifugation
    Authors:  Vamseedhar Rayaprolu, Jolene Ramsey, Joseph Che-Yen Wang and Suchetana Mukhopadhyay, date: 03/20/2018, view: 1498, Q&A: 0
    [Abstract] Chemical and sedimentation procedures are used to purify virus particles. While these approaches are successful for wild-type viruses, they are often not feasible for purifying mutant viruses with assembly defects. We combined two published methods (Atasheva et al., 2013; Moller-Tank et al., 2013), to generate a protocol that ...



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