Methods for Studying Mononuclear Phagocytes

Methods for Studying Mononuclear Phagocytes

1st Edition - December 28, 1981

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  • Editor: Dolph Adams
  • eBook ISBN: 9780323140690

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Methods for Studying Mononuclear Phagocytes is a practical guide to the study of mononuclear phagocytes that brings together various well-established and useful methods for examining these cells. The technical protocols have been made detailed, specific, practical, and inclusive of the necessary mystique for immediate and direct application in the laboratory. The book is divided into 11 parts arranged according to the sequence of steps that would generally be followed to study a given population of mononuclear phagocytes: (I) methods for obtaining and culturing populations of human and animal mononuclear phagocytes; (II) methods for separating populations of leukocytes to enrich or deplete their content of mononuclear phagocytes; (II) criteria and techniques for identifying mononuclear phagocytes; (IV) methods for quantifying the number of mononuclear phagocytes; (V) techniques for studying the morphology of mononuclear phagocytes; (VI) methods for quantifying the biochemical constituents of mononuclear phagocytes; (VII) methods of quantifying phagocytosis, pinocytosis, and chemotaxis; (VIII) methods for quantifying the secretory products of mononuclear phagocytes; (IX) procedures for quantifying the destruction of tumor cells and of microorganisms by mononuclear phagocytes; (X) methods for studying the cell biology of mononuclear phagocytes; and (XI) techniques for studying mononuclear phagocytes in vivo, including procedures for estimating their kinetics, accumulation, identification, and microbicidal properties.

Table of Contents

  • Contributors


    I. Obtaining and Culturing Mononuclear Phagocytes

    1 The Culture of Mononuclear Phagocytes: A Brief Overview

    2 Murine Mononuclear Phagocytes from Bone Marrow

    3 Obtaining and Culturing Murine Monocytes

    4 Obtaining and Culturing Human Monocytes

    5 Isolation of Human Monocytes

    6 Culture of Human Monocytes in Microplates and Enzymatic Assays for Following Their Maturation

    7 Peritoneal Mononuclear Phagocytes from Small Animals

    8 Obtaining and Culturing Human and Animal Alveolar Macrophages

    9 Obtaining Human Mononuclear Phagocytes from Colostrum

    10 Obtaining Adherent Cells from Spleen

    11 Obtaining Kupffer Cells

    12 Obtaining Mononuclear Phagocytes from Disaggregated Neoplasms

    13 Obtaining Mononuclear Phagocytes from Granulomas

    14 Teflon Film as a Substrate for the Culture of Mononuclear Phagocytes

    15 Effects of Husbandry and Mouse Strains on Mononuclear Phagocytes

    16 Endotoxin Contamination and in Vitro Monocyte-Macrophage Function: Methods of Detecting, Detoxifying, and Eliminating Endotoxin

    17 Continuous Cell Lines with Properties of Mononuclear Phagocytes

    II. Separation of Mononuclear Phagocytes for Enrichment or Depletion

    18 Separation of Mononuclear Phagocytes for Enrichment and for Depletion: An Overview

    19 Separation of Murine Macrophages by Adherence to Solid Substrates

    20 Separation of Human Monocytes and Guinea Pig Macrophages by Density Gradients of Metrizamide

    21 Separation of Murine Mononuclear Phagocytes by Density Gradients of Percoll

    22 Separation of Human Monocytes by Density Gradients of Percoll

    23 Separation of Murine Mononuclear Phagocytes by Velocity Sedimentation at Unit Gravity

    24 Use of Lidocaine for Detachment of Adherent Mononuclear Phagocytes

    25 Use of Sephadex Columns to Deplete Mononuclear Phagocytes

    26 Depletion of Mononuclear Phagocytes: Pitfalls in the Use of Carbonyl Iron, Carrageenan, Silica, Trypan Blue, or Anti-Mononuclear Phagocytes Serum

    III. Identification of Mononuclear Phagocytes

    27 Identification of Mononuclear Phagocytes: Overview and Definitions

    28 Characteristics of Mononuclear Phagocytes from Different Tissues

    29 Fc and C3 Receptors

    30 Identification of Mononuclear Phagocytes by Ingestion of Particulate Materials, such as Erythrocytes, Carbon, Zymosan, or Latex

    31 Heteroantisera Raised against Mononuclear Phagocytes

    32 Monoclonal Antibodies as Tools for the Study of Mononuclear Phagocytes

    33 Antisera against Ia Antigens

    IV. Quantitation of Number of Mononuclear Phagocytes

    34 Quantitation of Adherent Mononuclear Phagocytes by Inverted Phase Microscopy

    35 Quantitation of DNA in Mononuclear Phagocytes

    36 Lowry and Bradford Assays for Protein

    V. Morphology of Mononuclear Phagocytes

    37 Use of Wrights Strain and Cytocentrifuge Preparations

    38 Use of Phase Contrast Microscopy

    39 Use of Peroxidase Stain by the Kaplow Method

    40 Use of Nonspecific Esterase Stain

    41 Histochemical Stains for Macrophages in Cell Smears and Tissue Sections: β-Galactosidase, Acid Phosphatase, Nonspecific Esterase, Succinic Dehydrogenase, and Cytochrome Oxidase

    42 Use of Transmission Electron Microscopy

    43 Preparative Techniques for Scanning Electron Microscopy

    44 Use of Ultrastructural Histochemistry

    VI. Biochemical Constituents of Mononuclear Phagocytes

    45 Lysosomal Enzymes

    46 Microsomal Heme Oxygenase

    47 Histamine O-Methyltransferase

    48 5'-Nucleotidase Assay

    49 Alkaline Phosphodiesterase I

    50 Quantitation of Leucine Aminopeptidase of Mononuclear Phagocytes

    51 Hexose Monophosphate Shunt Activity and Oxygen Uptake

    52 Secretion of Superoxide Anion

    53 Release of Hydrogen Peroxide

    VII. General Functions of Mononuclear Phagocytes

    54 Antibody-Dependent and Antibody-Independent Phagocytosis

    55 Pinocytic Rate Using Horseradish Peroxidase

    56 Chemotaxis of Human and Murine Mononuclear Phagocytes

    VIII. Secretion by Mononuclear Phagocytes

    57 Secretory Functions of Mononuclear Phagocytes: Overview and Methods for Preparing Conditioned Supernatants

    58 Characterization and Classification of Macrophage Proteinases and Proteinase Inhibitors

    59 Growth of Macrophage on Collagen, Elastin, and Glycoprotein-Coated Plates as a Tool for Investigating Macrophage Proteinases

    60 Neutral Proteases by 3H-Labeled Casein

    61 Plasminogen Activators by Use of 3H-Labeled Casein Substrate

    62 Elastinolytic Enzymes

    63 Microtiter Assay for Antiviral Effects of Human and Murine Interferon Utilizing a Vertical Light Path Photometer for Quantitation

    64 Endogenous Pyrogen

    65 Prostaglandins

    66 Quantitation of Selected Complement Components

    67 Lysozyme

    IX. Destruction by Mononuclear Phagocytes

    68 Destruction of Listeria monocytogenes in Vitro

    69 Ingestion and Destruction of Candida albicans

    70 Quantitation of Destruction of Toxoplasma

    71 Destruction of Rickettsiae

    72 Destruction of Leishmania

    73 Destruction of Viruses

    74 Cytostasis of Tumor and Nontumor Cells

    75 Cytolysis of Tumor Cells by Release of [3H] Thymidine

    76 Quantification of Cytolysis of Neoplastic Cells by Release of Chromium-51

    77 Assessment of Cytolysis of Tumor Cells by Release of [125I] Iododeoxyuridine

    78 Antibody-Dependent Cellular Cytotoxicity (ADCC) of Erythroid and Tumor Cells

    X. Mononuclear Phagocytes as Tools in Cell Biology

    79 Overview: The Macrophage in Cell Biology

    80 Biosynthetic Radiolabeling of Cellular and Secreted Proteins of Mononuclear Phagocytes

    81 Extraction, Identification, and Quantitation of Lipids

    82 Synthesis, Cellular Turnover, and Mass of Cholesterol

    83 Solute Uptake and Membrane Transport by Mononuclear Phagocytes

    84 Use of Lactoperoxidase for Labeling Membrane Proteins

    85 Binding of Synthetic Chemotactic Peptides as a Model of Ligand-Receptor Interaction

    XI. Mononuclear Phagocytes In Vivo

    86 Isolation of Phagosomes from Mouse Peritoneal Macrophages

    87 Study of Mononuclear Macrophages in Vivo: Overview

    88 Quantitation of the Inflammatory Accumulation of Mononuclear Phagocytes in Vivo

    89 Systemic Labeling of Mononuclear Phagocytes

    90 Labeling of Mononuclear Phagocytes in Granulomas and Inflammatory Sites

    91 Identification of Fc and Complement Receptors in Tissue Sections

    92 Determination of Macrophage-Mediated Antibacterial Resistance

    Supplement to Table of Contents


Product details

  • No. of pages: 1048
  • Language: English
  • Copyright: © Academic Press 1981
  • Published: December 28, 1981
  • Imprint: Academic Press
  • eBook ISBN: 9780323140690

About the Editor

Dolph Adams

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