| Figures |
| Color Plates |
| Preface |
| Abbreviations |
| Introduction / 1： |
| Background |
| Advantages of Tissue Culture |
| Limitations |
| Major Differences In Vitro |
| Types of Tissue Culture |
| Biology of Cultured Cells / 2： |
| The Culture Environment |
| Cell Adhesion |
| Cell Proliferation |
| Differentiation |
| Energy Metabolism |
| Initiation of the Culture |
| Evolution of Cell Lines |
| The Development of Continuous Cell Lines |
| Origin of Cultured Cells |
| Design and Layout / 3： |
| Planning |
| Construction and Services |
| Layout |
| Equipment / 4： |
| Requirements of a Tissue Culture Laboratory Essential Equipment |
| Beneficial Equipment |
| Useful Additional Equipment |
| Consumable Items |
| Aseptic Technique / 5： |
| Objectives of Aseptic Technique |
| Elements of Aseptic Environment |
| Sterile Handling |
| Laminar Flow |
| Standard Procedure |
| Apparatus and Equipment |
| Safety / 6： |
| Risk Assessment |
| General Safety |
| Fire |
| Radiation |
| Biohazards |
| Culture Vessels / 7： |
| The Substrate |
| Choice of Culture Vessel |
| Specialized Systems |
| Treated Surfaces |
| Media / 8： |
| Development of Media |
| Physicochemical Properties |
| Balanced Salt Solutions |
| Complete Media |
| Serum |
| Selection of Medium and Serum |
| Other Supplements |
| Serum-Free Media / 9： |
| Disadvantages of Serum |
| Advantages of Serum-Free Media |
| Disadvantages of Serum-Free Media |
| Replacement of Serum |
| Selection of Serum-Free Medium |
| Development of Serum-Free Medium |
| Preparation of Serum-Free Medium |
| Conclusions |
| Preparation and Sterilization / 10： |
| Apparatus |
| Reagents and Media |
| Control, Testing, and Storage of Media |
| Primary Culture / 11： |
| Types of Primary Cell Culture |
| Isolation of the Tissue |
| Cell Lines / 12： |
| Nomenclature |
| Subculture and Propagation |
| Immortalization of Cell Lines |
| Cell Line Designations |
| Selection of Cell Line |
| Routine Maintenance |
| Cloning and Selection / 13： |
| Cloning |
| Stimulation of Plating Efficiency |
| Suspension Cloning |
| Isolation of Clones |
| Replica Plating |
| Selective Inhibitors |
| Isolation of Genetic Variants |
| Interaction with Susbstrate |
| Cell Separation / 14： |
| Cell Density and Isopyknic Sedimentation |
| Antibody-Based Techniques |
| Magnetic Sorting |
| Cell Size and Sedimentation Velocity |
| Centrifugal Elutriation |
| Fluorescence-Activated Cell Sorting |
| Other Techniques |
| Beginner's Approach to Cell Separation |
| Characterization / 15： |
| The Need for Characterization |
| Morphology |
| Chromosome Analysis |
| DNA Content |
| RNA and Protein |
| Enzyme Activity |
| Antigenic Markers |
| Authentication |
| Expression of In Vivo Phenotype / 16： |
| Stages of Commitment and Differentiation |
| Proliferation and Differentiation |
| Commitment and Lineage |
| Markers of Differentiation |
| Induction of Differentiation |
| Differentiation and Malignancy |
| Practical Aspects |
| Transformation / 17： |
| Role in Cell Line Characterization |
| What is Transformation? |
| Genetic Instability |
| Immortalization |
| Aberrant Growth Control |
| Tumorigenicity |
| Contamination / 18： |
| Sources of Contamination |
| Types of Microbial Contamination |
| Monitoring for Contamination |
| Eradication of Contamination |
| Cross-Contamination |
| Cryopreservation / 19： |
| Need for Cryopreservation |
| Preservation |
| Cell Banks |
| Transporting Cells |
| Quantitation / 20： |
| Cell Counting |
| Cell Weight |
| Protein |
| Rates of Synthesis |
| Preparation of Samples for Enzyme Assay and Immunoassay |
| Cytometry |
| Replicate Sampling |
| Plating Efficiency |
| Labeling Index |
| Cell Cycle Time (Generation Time) |
| Cell Migration |
| Cytotoxicity / 21： |
| In Vitro Limitations |
| Nature of the Assay |
| Anticancer Drug Screening |
| Inflammation |
| Specialized Cells / 22： |
| Epithelial Cells |
| Mensenchymal Cells |
| Neuroectodermal Cells |
| Hematopoietic Cells |
| Gonads |
| Tumor Cells / 23： |
| Sampling |
| Disaggregation |
| Development of Cell Lines |
| Selective Culture |
| Specific Tumor Types |
| Organotypic Culture / 24： |
| Cell Interaction and Phenotypic Expression |
| Organ Culture |
| Histotypic Culture |
| Filter-Well Inserts |
| Cultures of Neuronal Aggregates |
| Scale-Up / 25： |
| Scale-up in Suspension |
| Scale-up in Monolayer |
| Specialized Techniques / 26： |
| Lymphocyte Preparation |
| Autoradiography |
| Time-Lapse Recording |
| Confocal Microscopy |
| Cell Synchrony |
| Culture of Amniocytes |
| Culture of Cells from Poikilotherms |
| Molecular Techniques / 27： |
| Molecular Biology in Cell Culture |
| In Situ Molecular Hybridization |
| Somatic Cell Fusion |
| Production of Monoclonal Antibodies |
| DNA Transfer |
| Problem Solving / 28： |
| Slow Cell Growth |
| Choice of Medium |
| Unstable Reagents |
| Purity of Constituents |
| Plastics |
| Glassware |
| Microbial Contamination |
| Chemical Contamination |
| Feeding |
| Subculture |
| Granularity of Cells |
| Viability |
| In Conclusion |
| Reagent Appendix |
| Trade Index |
| Glossary |
| References |
| Index |
