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| Figures | |
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| Color Plates | |
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| Preface | |
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| Abbreviations | |
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| Introduction | |
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| Background | |
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| Advantages of Tissue Culture | |
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| Limitations | |
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| Major Differences In Vitro | |
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| Types of Tissue Culture | |
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| Biology of Cultured Cells | |
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| The Culture Environment | |
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| Cell Adhesion | |
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| Cell Proliferation | |
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| Differentiation | |
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| Energy Metabolism | |
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| Initiation of the Culture | |
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| Evolution of Cell Lines | |
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| The Development of Continuous Cell Lines | |
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| Origin of Cultured Cells | |
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| Design and Layout | |
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| Planning | |
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| Construction and Services | |
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| Layout | |
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| Equipment | |
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| Requirements of a Tissue Culture Laboratory Essential Equipment | |
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| Beneficial Equipment | |
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| Useful Additional Equipment | |
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| Consumable Items | |
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| Aseptic Technique | |
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| Objectives of Aseptic Technique | |
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| Elements of Aseptic Environment | |
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| Sterile Handling | |
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| Laminar Flow | |
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| Standard Procedure | |
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| Apparatus and Equipment | |
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| Safety | |
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| Risk Assessment | |
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| General Safety | |
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| Fire | |
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| Radiation | |
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| Biohazards | |
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| Culture Vessels | |
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| The Substrate | |
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| Choice of Culture Vessel | |
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| Specialized Systems | |
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| Treated Surfaces | |
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| Media | |
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| Development of Media | |
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| Physicochemical Properties | |
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| Balanced Salt Solutions | |
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| Complete Media | |
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| Serum | |
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| Selection of Medium and Serum | |
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| Other Supplements | |
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| Serum-Free Media | |
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| Disadvantages of Serum | |
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| Advantages of Serum-Free Media | |
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| Disadvantages of Serum-Free Media | |
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| Replacement of Serum | |
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| Selection of Serum-Free Medium | |
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| Development of Serum-Free Medium | |
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| Preparation of Serum-Free Medium | |
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| Conclusions | |
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| Preparation and Sterilization | |
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| Apparatus | |
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| Reagents and Media | |
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| Control, Testing, and Storage of Media | |
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| Primary Culture | |
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| Types of Primary Cell Culture | |
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| Isolation of the Tissue | |
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| Primary Culture | |
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| Cell Lines | |
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| Nomenclature | |
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| Subculture and Propagation | |
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| Immortalization of Cell Lines | |
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| Cell Line Designations | |
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| Selection of Cell Line | |
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| Routine Maintenance | |
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| Cloning and Selection | |
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| Cloning | |
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| Stimulation of Plating Efficiency | |
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| Suspension Cloning | |
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| Isolation of Clones | |
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| Replica Plating | |
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| Selective Inhibitors | |
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| Isolation of Genetic Variants | |
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| Interaction with Susbstrate | |
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| Cell Separation | |
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| Cell Density and Isopyknic Sedimentation | |
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| Antibody-Based Techniques | |
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| Magnetic Sorting | |
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| Cell Size and Sedimentation Velocity | |
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| Centrifugal Elutriation | |
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| Fluorescence-Activated Cell Sorting | |
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| Other Techniques | |
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| Beginner's Approach to Cell Separation | |
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| Characterization | |
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| The Need for Characterization | |
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| Morphology | |
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| Chromosome Analysis | |
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| DNA Content | |
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| RNA and Protein | |
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| Enzyme Activity | |
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| Antigenic Markers | |
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| Differentiation | |
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| Authentication | |
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| Differentiation | |
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| Expression of In Vivo Phenotype | |
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| Stages of Commitment and Differentiation | |
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| Proliferation and Differentiation | |
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| Commitment and Lineage | |
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| Markers of Differentiation | |
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| Induction of Differentiation | |
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| Differentiation and Malignancy | |
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| Practical Aspects | |
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| Transformation | |
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| Role in Cell Line Characterization | |
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| What is Transformation? | |
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| Genetic Instability | |
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| Immortalization | |
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| Aberrant Growth Control | |
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| Tumorigenicity | |
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| Contamination | |
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| Sources of Contamination | |
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| Types of Microbial Contamination | |
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| Monitoring for Contamination | |
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| Eradication of Contamination | |
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| Cross-Contamination | |
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| Conclusions | |
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| Cryopreservation | |
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| Need for Cryopreservation | |
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| Preservation | |
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| Cell Banks | |
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| Transporting Cells | |
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| Quantitation | |
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| Cell Counting | |
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| Cell Weight | |
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| DNA Content | |
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| Protein | |
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| Rates of Synthesis | |
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| Preparation of Samples for Enzyme Assay and Immunoassay | |
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| Cytometry | |
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| Replicate Sampling | |
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| Cell Proliferation | |
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| Plating Efficiency | |
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| Labeling Index | |
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| Cell Cycle Time (Generation Time) | |
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| Cell Migration | |
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| Cytotoxicity | |
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| Introduction | |
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| In Vitro Limitations | |
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| Nature of the Assay | |
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| Anticancer Drug Screening | |
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| Transformation | |
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| Inflammation | |
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| Specialized Cells | |
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| Epithelial Cells | |
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| Mensenchymal Cells | |
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| Neuroectodermal Cells | |
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| Hematopoietic Cells | |
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| Gonads | |
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| Tumor Cells | |
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| Sampling | |
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| Disaggregation | |
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| Primary Culture | |
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| Characterization | |
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| Development of Cell Lines | |
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| Selective Culture | |
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| Specific Tumor Types | |
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| Organotypic Culture | |
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| Cell Interaction and Phenotypic Expression | |
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| Organ Culture | |
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| Histotypic Culture | |
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| Filter-Well Inserts | |
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| Cultures of Neuronal Aggregates | |
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| Organotypic Culture | |
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| Scale-Up | |
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| Scale-up in Suspension | |
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| Scale-up in Monolayer | |
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| Specialized Techniques | |
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| Lymphocyte Preparation | |
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| Autoradiography | |
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| Time-Lapse Recording | |
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| Confocal Microscopy | |
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| Cell Synchrony | |
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| Culture of Amniocytes | |
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| Culture of Cells from Poikilotherms | |
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| Molecular Techniques | |
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| Molecular Biology in Cell Culture | |
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| In Situ Molecular Hybridization | |
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| Somatic Cell Fusion | |
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| Production of Monoclonal Antibodies | |
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| DNA Transfer | |
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| Problem Solving | |
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| Slow Cell Growth | |
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| Choice of Medium | |
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| Unstable Reagents | |
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| Purity of Constituents | |
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| Plastics | |
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| Glassware | |
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| Microbial Contamination | |
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| Chemical Contamination | |
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| Primary Culture | |
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| Cloning | |
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| Differentiation | |
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| Feeding | |
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| Subculture | |
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| Cross-Contamination | |
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| Cryopreservation | |
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| Granularity of Cells | |
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| Cell Counting | |
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| Viability | |
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| In Conclusion | |
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| Reagent Appendix | |
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| Trade Index | |
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| Glossary | |
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| References | |
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| Index | |