Cancer Biology

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    Protocols in Current Issue
    Generation of the Compression-induced Dedifferentiated Adipocytes (CiDAs) Using Hypertonic Medium
    [Abstract]

    Current methods to obtain mesenchymal stem cells (MSCs) involve sampling, culturing, and expanding of primary MSCs from adipose, bone marrow, and umbilical cord tissues. However, the drawbacks are the limited numbers of total cells in MSC pools, and their decaying stemness during in vitro expansion. As an alternative resource, recent ceiling

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    Carboxyfluorescein Dye Uptake to Measure Connexin-mediated Hemichannel Activity in Cultured Cells
    [Abstract]

    Connexins are membrane bound proteins that facilitate direct and local paracrine mediated cell-to-cell communication through their ability to oligomerise into hexameric hemichannels. When neighbouring channels align, they form gap-junctions that provide a direct route for information transfer between cells. In contrast to intact gap junctions,

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    A Transient Transfection-based Cell Adhesion Assay with 293T Cells
    Authors:  Rohit Singh and Beom K. Choi, date: 01/05/2021, view: 766, Q&A: 0
    [Abstract]

    The in vitro cell adhesion assay is a quantitative method for measuring selective cell adhesion to specific proteins. Traditionally, cell adhesion assays employ purified protein immobilized on a solid glass or plastic surface. Here, we describe a transient 293T cell transfection-based cell adhesion assay to study selective cell adhesion of a

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    A Rigorous Quantitative Approach to Analyzing Phagocytosis Assays
    Authors:  Michael D. Caponegro, Kaitlyn Koenig Thompson, Maryam Tayyab and Stella E. Tsirka, date: 01/05/2021, view: 1578, Q&A: 0
    [Abstract] Studying monocytic cells in isolated systems in vitro contributes significantly to the understanding of innate immune physiology. Functional assays produce read outs which can be used to measure responses to selected stimuli, such as pathogen exposure, antigen loading, and cytokine stimulation. Integration of these results with high ...
    Triacylglycerol Measurement in HeLa Cells
    Authors:  Ximing Du and Hongyuan Yang, date: 12/20/2020, view: 893, Q&A: 0
    [Abstract]

    Lipid droplets store triacylglycerols (triglycerides) and sterol esters to regulate lipid and energy homeostasis. Triacylglycerol measurement is often performed during the investigation of lipid droplet formation and growth. This protocol describes a reliable method using a fluorometric lipid quantification kit to measure triacylglycerols

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    Automated Analysis of Cell Surface Ruffling: Ruffle Quantification Macro
    Authors:  Nicholas D. Condon, Jennifer L. Stow and Adam A. Wall, date: 01/20/2020, view: 1854, Q&A: 1
    [Abstract] Cell surface protrusions include F-actin rich, wave-like ruffles that are erected transiently in response to stimuli and during cell migration. Macrophages are innate immune cells that ruffle constitutively and more dramatically in cells activated by pathogens. Dorsal ruffles and their resulting macropinosomes are key sites for environmental ...
    Immunofluorescence-based Determination of Centrosome Number in Tissue Samples
    Authors:  Mengdie Wang, Gregory C. Rogers and Anne E. Cress, date: 10/20/2019, view: 2249, Q&A: 0
    [Abstract] Centrosome numerical abnormalities have been reported in a variety of tumors. Centrosome numbers in cancer cells display both inter-tumor and intra-tumor heterogeneity. The over production of centrosomes (centrosome amplification) is unique in cancer cells and is a promising target for therapy. Thus, a method to quantify centrosome numbers on a ...
    RNA Interference Screening to Identify Proliferation Determinants in Breast Cancer Cells
    [Abstract] RNAi screening technology has revealed unknown determinants of various biological signaling pathways in biomedical studies. This protocol provided detailed information about how to use RNAi screening to identify proliferation determinants in breast tumor cells. siRNA-based libraries targeting against Estrogen receptor (ER)-network, including 631 ...
    Measurement of the Intracellular Calcium Concentration with Fura-2 AM Using a Fluorescence Plate Reader
    Authors:  Magdiel Martínez, Namyr A. Martínez and Walter I. Silva, date: 07/20/2017, view: 24265, Q&A: 1
    [Abstract] Intracellular calcium elevation triggers a wide range of cellular responses. Calcium responses can be affected or modulated by membrane receptors mutations, localization, exposure to agonists/antagonists, among others (Burgos et al., 2007; Martínez et al., 2016). Changes in intracellular calcium concentration can be measured ...
    Soft Agar Colony Formation Assay as a Hallmark of Carcinogenesis
    Authors:  Feng Du, Xiaodi Zhao and Daiming Fan, date: 06/20/2017, view: 22145, Q&A: 0
    [Abstract] Soft agar colony formation assay is established to estimate the anchorage-independent growth ability of cells. In this assay, a bottom layer of agar with complete media is poured and solidified first, followed by an upper layer containing a specified number of cells suspended in medium-agar mixture. After two weeks of incubation, the number of ...



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