Cell Biology

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    Protocols in Current Issue
    Observing Nutrient Gradients, Gene Expression and Growth Variation Using the "Yeast Machine" Microfluidic Device
    [Abstract] The natural environment of microbial cells like bacteria and yeast is often a complex community in which growth and internal organization reflect morphogenetic processes and interactions that are dependent on spatial position and time. While most of research is performed in simple homogeneous environments (e.g., bulk liquid cultures), ...
    Isolation of Tumor Cells Based on Their Distance from Blood Vessels
    Authors:  Saran Kumar, Husni Sharife, Tirzah Kreisel, Libat Bar-Lev, Myriam Grunewald and Eli Keshet, date: 05/20/2020, view: 1165, Q&A: 0
    [Abstract] Differential exposure of tumor cells to microenvironmental cues greatly impacts cell phenotypes, raising a need for position based sorting of tumor cells amenable to multiple OMICs and functional analyses. One such key determinant of tumor heterogeneity in solid tumors is its vasculature. Proximity to blood vessels (BVs) profoundly affects tumor ...
    Steady-state and Flux-based Trehalose Estimation as an Indicator of Carbon Flow from Gluconeogenesis or Glycolysis
    Authors:  Ritu Gupta and Sunil Laxman, date: 01/05/2020, view: 1369, Q&A: 0
    [Abstract] Trehalose (and glycogen) is a major storage carbohydrate in many cells, including S. cerevisiae. Typically, trehalose (a disaccharide of glucose) is synthesized and stored through gluconeogenesis. However, trehalose can also be made directly from glucose, if glucose-6-phosphate is channeled away from glycolysis or pentose phosphate ...
    Determination of the Glycolysis and Lipogenesis in Culture of Hepatocytes
    Authors:  Pierre-Damien Denechaud and Luis Fajas, date: 11/05/2016, view: 8511, Q&A: 0
    [Abstract] Metabolic flux analyses are needed to provide insights into metabolic regulation that occurs in cells. The current protocol describes fast and reproducible methods for determining glycolysis and de novo lipogenesis of hepatocytes. Primary culture of hepatocytes is an ‘in vitro’ model useful to study liver glucose and lipid ...
    Stable Isotope Resolved Metabolomics Studies in ex vivo TIssue Slices
    Authors:  Teresa W-M. Fan, Andrew N. Lane and Richard M. Higashi, date: 02/05/2016, view: 7162, Q&A: 0
    [Abstract] An important component of this methodology is to assess the role of the tumor microenvironment on tumor growth and survival. To tackle this problem, we have adapted the original approach of Warburg (Warburg, 1923), by combining thin tissue slices with Stable Isotope Resolved Metabolomics (SIRM) to determine detailed metabolic activity of human ...
    13C Tracer Studies of Metabolism in Mouse Tumor Xenografts
    Authors:  Andrew N. Lane, Jun Yan and Teresa W-M. Fan, date: 11/20/2015, view: 8300, Q&A: 0
    [Abstract] Mice are widely used for human tumor xenograft studies of cancer development and drug efficacy and toxicity. Stable isotope tracing coupled with metabolomic analysis is an emerging approach for assaying metabolic network activity. In mouse models there are several routes of tracer introduction, which have particular advantages and disadvantages ...
    Assessment of Brown Adipocyte Thermogenic Function by High-throughput Respirometry
    Authors:  Kiana Mahdaviani, Ilan Benador and Orian Shirihai, date: 11/05/2015, view: 10417, Q&A: 0
    [Abstract] Brown adipose tissue (BAT) has the unique ability to dramatically increase mitochondrial uncoupled fuel oxidation for thermogenesis in response to adrenergic stimulation. A key parameter in assessing brown adipocyte thermogenic capacity is mitochondrial uncoupling as determined by respiration. Measuring mitochondrial oxygen consumption rate (OCR) ...
    ATP and Lactate Quantification
    Authors:  Isaline Rowe, Marco Chiaravalli and Alessandra Boletta, date: 12/05/2013, view: 12894, Q&A: 0
    [Abstract] Cells use glucose to generate energy by two different metabolic processes: lactic fermentation and aerobic respiration. In the first common series of reactions, glucose is converted into pyruvate. In anaerobic conditions, pyruvate is transformed into lactate, this process yields to 2 ATP molecules per glucose molecule. In the presence of oxygen, ...
    Detection of Released CO2 by Radioactive Lactate
    Authors:  Tania Fiaschi and Paola Chiarugi, date: 07/05/2013, view: 6560, Q&A: 0
    [Abstract] This method allows to evaluate the degradation of lactate during cellular respiration. During this metabolic process, carbon atoms of lactate can be transformed in carbon dioxide. For this purpose, the radioactive lactate is added to the cells and the amount of radioactive carbon dioxide liberated is monitored. The radioactive carbon dioxide ...
    Analysis of the Incorporation of Carbon Atoms from Radioactive Lactate into Proteins
    Authors:  Tania Fiaschi and Paola Chiarugi, date: 07/05/2013, view: 5612, Q&A: 0
    [Abstract] This method allows to analyze if the carbon atoms of lactate are embedded into proteins. Indeed, mammalian cells express the transporter of monocarboxylic acids (called MCT1) that allows the entry of lactate into the cell. To this end, cells are incubated for 24 h with the culture medium containing lactate uniformly labeled with carbon 14 and ...



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