Cell Biology

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    Protocols in Current Issue
    A High-throughput and Pathophysiologically Relevant Astrocyte-motor Neuron Co-culture Assay for Amyotrophic Lateral Sclerosis Therapeutic Discovery
    Authors:  Matthew J. Stopford, Scott P. Allen and Laura Ferraiuolo, date: 09/05/2019, view: 396, Q&A: 0
    Amyotrophic lateral sclerosis (ALS) is an adult onset neurological disorder characterized by loss of motor neurons leading to progressive muscle wasting and eventually death. Astrocytes play a key role in disease pathogenesis. However, the ability ...
    In vitro Intestinal Epithelial Wound-healing Assays Using Electric Cell-Substrate Impedance Sensing Instrument
    Authors:  Olivier Merlin-Zhang, Junsik Sung and Emilie Viennois, date: 09/05/2019, view: 205, Q&A: 0
    Here, we describe an in vitro epithelial wound-healing assay using Electric Cell-Substrate Impedance Sensing (ECIS) technology. The ECIS technology is a real time cell growth assay based on a small (250 μm diameter) active gold electrode ...
    Co-culture of Human Stem Cell Derived Neurons and Oligodendrocyte Progenitor Cells
    Authors:  Stephanie Dooves, Aishwarya G. Nadadhur, Lisa Gasparotto and Vivi M. Heine, date: 09/05/2019, view: 285, Q&A: 0
    Crosstalk between neurons and oligodendrocytes is important for proper brain functioning. Multiple co-culture methods have been developed to study oligodendrocyte maturation, myelination or the effect of oligodendrocytes on neurons. However, most of ...
    High Resolution Respirometry in Candida albicans
    Authors:  Lucian Duvenage, Carol A. Munro and Campbell W. Gourlay, date: 09/05/2019, view: 141, Q&A: 0
    Many Candida species, such as the opportunistic human pathogen Candida albicans, are Crabtree-Negative yeasts and are therefore highly dependent on the energy generated through oxidative phosphorylation. Respiration control is ...
    A Widely Applicable Urea-based Fluorescent/Colorimetric mRNA in situ Hybridization Protocol
    Author:  Chiara Sinigaglia, date: 09/05/2019, view: 235, Q&A: 0
    In situ hybridization methods are routinely employed to detect nucleic acid sequences, allowing to localize gene expression or to study chromosomal organization in their native context. These methods rely on the pairwise binding of a ...
    Isolation of Neutrophil Nuclei for Use in NETosis Assays
    Author:  Dave Boucher, date: 09/05/2019, view: 265, Q&A: 0
    Neutrophils are critical immune cells that protect our body against invading pathogens. They generate antibacterial DNA structures called neutrophil extracellular traps (NET). Recently we identified a new mechanism that enables NET formation. We ...
    Estrous Cycle Monitoring in Mice with Rapid Data Visualization and Analysis
    Authors:  Leanna K. Pantier, Jiang Li and Catherine A. Christian, date: 09/05/2019, view: 316, Q&A: 0
    The estrous cycle provides a readout of reproductive health in female laboratory rodents, and estrous cycle stage can be an important physiological variable. Accurate assessment of estrous cycle stage is also important in producing timed pregnancies ...
    A High-throughput and Pathophysiologically Relevant Astrocyte-motor Neuron Co-culture Assay for Amyotrophic Lateral Sclerosis Therapeutic Discovery
    Authors:  Matthew J. Stopford, Scott P. Allen and Laura Ferraiuolo, date: 09/05/2019, view: 396, Q&A: 0
    [Abstract] Amyotrophic lateral sclerosis (ALS) is an adult onset neurological disorder characterized by loss of motor neurons leading to progressive muscle wasting and eventually death. Astrocytes play a key role in disease pathogenesis. However, the ability to study astrocytic support towards motor neurons in ALS has been limited by a lack of sustainable ...
    In vitro Intestinal Epithelial Wound-healing Assays Using Electric Cell-Substrate Impedance Sensing Instrument
    Authors:  Olivier Merlin-Zhang, Junsik Sung and Emilie Viennois, date: 09/05/2019, view: 205, Q&A: 0
    [Abstract] Here, we describe an in vitro epithelial wound-healing assay using Electric Cell-Substrate Impedance Sensing (ECIS) technology. The ECIS technology is a real time cell growth assay based on a small (250 μm diameter) active gold electrode which resistance is measured continuously. When intestinal epithelial cells reach confluency on the ...
    Co-culture of Human Stem Cell Derived Neurons and Oligodendrocyte Progenitor Cells
    Authors:  Stephanie Dooves, Aishwarya G. Nadadhur, Lisa Gasparotto and Vivi M. Heine, date: 09/05/2019, view: 285, Q&A: 0
    [Abstract] Crosstalk between neurons and oligodendrocytes is important for proper brain functioning. Multiple co-culture methods have been developed to study oligodendrocyte maturation, myelination or the effect of oligodendrocytes on neurons. However, most of these methods contain cells derived from animal models. In the current protocol, we co-culture ...
    High Resolution Respirometry in Candida albicans
    Authors:  Lucian Duvenage, Carol A. Munro and Campbell W. Gourlay, date: 09/05/2019, view: 141, Q&A: 0
    [Abstract] Many Candida species, such as the opportunistic human pathogen Candida albicans, are Crabtree-Negative yeasts and are therefore highly dependent on the energy generated through oxidative phosphorylation. Respiration control is linked to a range of aspects of C. albicans cell physiology that appear to be important for ...
    A Widely Applicable Urea-based Fluorescent/Colorimetric mRNA in situ Hybridization Protocol
    Author:  Chiara Sinigaglia, date: 09/05/2019, view: 235, Q&A: 0
    [Abstract] In situ hybridization methods are routinely employed to detect nucleic acid sequences, allowing to localize gene expression or to study chromosomal organization in their native context. These methods rely on the pairwise binding of a labeled probe to the target endogenous nucleic acid sequence–the hybridization step, followed by detection ...
    Isolation of Neutrophil Nuclei for Use in NETosis Assays
    Author:  Dave Boucher, date: 09/05/2019, view: 265, Q&A: 0
    [Abstract] Neutrophils are critical immune cells that protect our body against invading pathogens. They generate antibacterial DNA structures called neutrophil extracellular traps (NET). Recently we identified a new mechanism that enables NET formation. We observed that following recognition of lipopolysaccharides, inflammatory caspases cleave Gasdermin D ...
    Estrous Cycle Monitoring in Mice with Rapid Data Visualization and Analysis
    Authors:  Leanna K. Pantier, Jiang Li and Catherine A. Christian, date: 09/05/2019, view: 316, Q&A: 0
    [Abstract] The estrous cycle provides a readout of reproductive health in female laboratory rodents, and estrous cycle stage can be an important physiological variable. Accurate assessment of estrous cycle stage is also important in producing timed pregnancies for developmental studies. Here, we provide a protocol for evaluation of estrous cycle stage ...
    In vitro Differentiation of Thymic Treg Cell Progenitors to Mature Thymic Treg Cells
    Authors:  David L. Owen and Michael A. Farrar, date: 08/20/2019, view: 217, Q&A: 0
    [Abstract] Thymic Treg cell differentiation occurs via a two-step process. Step one generates Treg cell progenitors (TregP) and is driven by strong TCR interactions with antigens presented in the thymus. Step two is initiated by activation of STAT5 via IL-2, or IL-15, leading to mature Treg cells capable of ...
    Fibroblast Gap-closure Assay-Microscopy-based in vitro Assay Measuring the Migration of Murine Fibroblasts
    Authors:  Agnieszka P. Looney and Mallar Bhattacharya, date: 08/20/2019, view: 348, Q&A: 0
    [Abstract] Pulmonary fibrosis is characterized by pathological scaring of the lung. Similar to other fibrotic diseases, scar formation is driven by excessive extracellular matrix deposition by activated, proliferative, and migratory fibroblasts.

    Currently, the two most widely used chemotaxis and cell migration assays are the scratch assay and the ...
    Non-invasive Quantification of Cell Wall Porosity by Fluorescence Quenching Microscopy
    Authors:  Xiaohui Liu, Thomas Günther Pomorski and Johannes Liesche, date: 08/20/2019, view: 204, Q&A: 0
    [Abstract] All bacteria, fungi and plant cells are surrounded by a cell wall. This complex network of polysaccharides and glycoproteins provides mechanical support, defines cell shape, controls cell growth and influences the exchange of substances between the cell and its surroundings. Despite its name, the cell wall is a flexible, dynamic structure. ...



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