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Ian Freshney ; reviewing editors, Amanda Capes-Davis, Carl Gregory, Stefan Przyborski 205 $aSeventh edition. 210 1$aHoboken, New Jersey :$cWiley-Blackwell,$d2016. 210 4$d©2016 215 $a1 online resource (758 p.) 300 $aDescription based upon print version of record. 311 $a1-118-87365-3 320 $aIncludes bibliographical references at the end of each chapters and index. 327 $aIntro -- Titlepage -- Copyright -- Dedication -- List of Color Plates -- List of Protocols -- List of Minireviews -- Preface and Acknowledgments -- Abbreviations -- About the Companion Website -- CHAPTER 1 Introduction -- 1.1 HISTORICAL BACKGROUND -- 1.2 ADVANTAGES OF TISSUE CULTURE -- 1.3 LIMITATIONS -- 1.4 MAJOR DIFFERENCES IN VITRO -- 1.5 TYPES OF TISSUE CULTURE -- REFERENCES -- CHAPTER 2 Biology of Cultured Cells -- 2.1 THE CULTURE ENVIRONMENT -- 2.2 CELL ADHESION -- 2.3 CELL PROLIFERATION -- 2.4 DIFFERENTIATION -- 2.5 CELL SIGNALING -- 2.6 ENERGY METABOLISM -- 2.7 ORIGIN OF CULTURED CELLS -- 2.8 DEFINITIONS -- REFERENCES -- CHAPTER 3 Laboratory Design and Layout -- 3.1 PLANNING, FURNISHING, AND SERVICES -- 3.2 DESIGN AND LAYOUT -- 3.3 DISASTER MANAGEMENT -- REFERENCES -- CHAPTER 4 Equipment and Materials -- 4.1 REQUIREMENTS OF A TISSUE CULTURE LABORATORY -- 4.2 STERILE WORKING AREA -- 4.3 INCUBATION AND CULTURE -- 4.4 PREPARATION AND STERILIZATION -- 4.5 STORAGE -- 4.6 SUPPLEMENTARY LABORATORY EQUIPMENT -- 4.7 SPECIALIZED EQUIPMENT -- REFERENCES -- CHAPTER 5 Aseptic Technique -- 5.1 OBJECTIVES OF ASEPTIC TECHNIQUE -- 5.2 ELEMENTS OF ASEPTIC ENVIRONMENT -- 5.3 STERILE HANDLING -- 5.4 STANDARD PROCEDURE -- 5.5 APPARATUS AND EQUIPMENT -- REFERENCES -- CHAPTER 6 Safety, Bioethics, and Validation -- 6.1 LABORATORY SAFETY -- 6.2 RISK ASSESSMENT -- 6.3 STANDARD OPERATING PROCEDURES -- 6.4 SAFETY REGULATIONS -- 6.5 GENERAL SAFETY -- 6.6 FIRE -- 6.7 IONIZING RADIATION -- 6.8 BIOHAZARDS -- 6.9 BIOETHICS -- 6.10 QUALITY ASSURANCE -- REFERENCES -- CHAPTER 7 Culture Vessels and Substrates -- 7.1 THE SUBSTRATE -- 7.2 TREATED SURFACES -- 7.3 CHOICE OF CULTURE VESSEL -- 7.4 SPECIALIZED SYSTEMS -- REFERENCES -- CHAPTER 8 Defined Media and Supplements -- 8.1 DEVELOPMENT OF MEDIA -- 8.2 PHYSICOCHEMICAL PROPERTIES -- 8.3 BALANCED SALT SOLUTIONS. 327 $a8.4 COMPLETE MEDIA -- 8.5 SERUM -- 8.6 SELECTION OF MEDIUM AND SERUM -- 8.7 OTHER SUPPLEMENTS -- 8.8 STORAGE -- REFERENCES -- CHAPTER 9 Serum-Free Media -- 9.1 DISADVANTAGES OF SERUM -- 9.2 ADVANTAGES OF SERUM-FREE MEDIA -- 9.3 DISADVANTAGES OF SERUM-FREE MEDIA -- 9.4 REPLACEMENT OF SERUM -- 9.5 DEVELOPMENT OF SERUM-FREE MEDIUM -- 9.6 SELECTION OF SERUM-FREE MEDIUM -- 9.7 PREPARATION OF SERUM-FREE MEDIUM -- 9.8 ANIMAL PROTEIN?FREE MEDIA -- 9.9 CONCLUSIONS -- REFERENCES -- CHAPTER 10 Preparation and Sterilization -- 10.1 PREPARATION OF REAGENTS AND MATERIALS -- 10.2 STERILIZATION OF APPARATUS AND LIQUIDS -- 10.3 APPARATUS -- 10.4 REAGENTS AND MEDIA -- 10.5 STERILIZATION OF MEDIA -- 10.6 CONTROL, TESTING, AND STORAGE OF MEDIA -- REFERENCES -- CHAPTER 11 Primary Culture -- 11.1 INITIATION OF A PRIMARY CELL CULTURE -- 11.2 ISOLATION OF THE TISSUE -- 11.3 TYPES OF PRIMARY CULTURE -- REFERENCES -- CHAPTER 12 Subculture and Cell Lines -- 12.1 TERMINOLOGY -- 12.2 SUBCULTURE AND PROPAGATION -- 12.3 CHOOSING A CELL LINE -- 12.4 ROUTINE MAINTENANCE -- 12.5 SUBCULTURE -- REFERENCES -- CHAPTER 13 Authentication and Validation -- 13.1 AUTHENTICATION OF CELL LINES -- 13.2 VALIDATION -- 13.3 QUALITY ASSURANCE -- REFERENCES -- CHAPTER 14 Microbial Contamination -- 14.1 SOURCES OF CONTAMINATION -- 14.2 TYPES OF MICROBIAL CONTAMINATION -- 14.3 MONITORING FOR CONTAMINATION -- 14.4 DISPOSAL OF CONTAMINATED CULTURES -- 14.5 ERADICATION OF CONTAMINATION -- 14.6 CROSS-CONTAMINATION -- 14.7 CONCLUSIONS -- REFERENCES -- CHAPTER 15 Cryopreservation and Banking -- 15.1 RATIONALE FOR FREEZING -- 15.2 CONSIDERATIONS BEFORE CRYOPRESERVATION -- 15.3 PRINCIPLES OF CRYOPRESERVATION -- 15.4 DESIGN AND CONTROL OF FREEZER STOCKS -- 15.5 CELL BANKS -- 15.6 TRANSPORTING CELLS -- REFERENCES -- CHAPTER 16 Cloning and Selection -- 16.1 CELL CLONING -- 16.3 SUSPENSION CLONING. 327 $a16.4 ISOLATION OF CLONES -- 16.5 REPLICA PLATING -- 16.6 SELECTIVE MEDIA AND INHIBITORS -- 16.7 INTERACTION WITH SUBSTRATE -- REFERENCES -- CHAPTER 17 Cell Sorting -- 17.1 CELL DENSITY AND ISOPYKNIC SEDIMENTATION -- 17.2 CELL SIZE AND SEDIMENTATION VELOCITY -- 17.3 ANTIBODY-BASED TECHNIQUES -- 17.4 BEGINNER'S APPROACH TO CELL SEPARATION -- REFERENCES -- CHAPTER 18 Cell Line Characterization -- 18.1 THE NEED FOR CHARACTERIZATION -- 18.2 GENOTYPIC CHARACTERIZATION -- 18.3 PHENOTYPIC CHARACTERIZATION -- 18.4 CELL MORPHOLOGY -- 18.5 ANTIGENIC MARKERS -- 18.6 TIME-LAPSE RECORDING -- 18.7 CELL CYCLE -- REFERENCES -- CHAPTER 19 Differentiation -- 19.1 EXPRESSION OF THE IN VIVO PHENOTYPE -- 19.2 STAGES OF DIFFERENTIATION -- 19.3 PROLIFERATION AND DIFFERENTIATION -- 19.4 COMMITMENT AND LINEAGE -- 19.5 STEM CELL PLASTICITY -- 19.6 MARKERS OF DIFFERENTIATION -- 19.7 INDUCTION OF DIFFERENTIATION -- 19.8 DIFFERENTIATION AND MALIGNANCY -- 19.9 PRACTICAL ASPECTS -- REFERENCES -- CHAPTER 20 Three-Dimensional Culture -- 20.1 CELL INTERACTION AND PHENOTYPIC EXPRESSION -- 20.2 ORGAN CULTURE -- 20.3 HISTOTYPIC CULTURE -- 20.4 ORGANOTYPIC CULTURE -- 20.5 IMAGING CELLS IN 3D CONSTRUCTS -- REFERENCES -- CHAPTER 21 Scale-up and Automation -- 21.1 SCALE-UP IN SUSPENSION -- 21.2 SCALE-UP IN MONOLAYER -- 21.3 PROCESS CONTROL -- 21.4 LARGE-SCALE PROCESSES -- 21.5 AUTOMATION -- REFERENCES -- CHAPTER 22 Senescence, Immortalization, and Transformation -- 22.1 ROLE IN CELL LINE CHARACTERIZATION -- 22.2 GENETIC INSTABILITY AND HETEROGENEITY -- 22.3 IMMORTALIZATION -- 22.4 ABERRANT GROWTH CONTROL -- 22.5 TUMORIGENICITY -- REFERENCES -- CHAPTER 23 Quantitation -- 23.1 CELL COUNTING -- 23.2 CELL WEIGHT AND PACKED CELL VOLUME -- 23.3 DNA CONTENT -- 23.4 PROTEIN -- 23.5 RATES OF SYNTHESIS -- 23.6 PREPARATION OF SAMPLES FOR ENZYME ASSAY AND IMMUNOASSAY -- 23.7 CYTOMETRY. 327 $a23.8 REPLICATE SAMPLING -- 23.9 CELL PROLIFERATION -- 23.10 PLATING EFFICIENCY -- 23.11 LABELING INDEX -- 23.12 AUTORADIOGRAPHY -- 23.13 CELL CYCLE TIME -- 23.14 CELL MIGRATION -- REFERENCES -- CHAPTER 24 Cytotoxicity -- 24.1 VIABILITY, TOXICITY, AND SURVIVAL -- 24.2 IN VITRO LIMITATIONS -- 24.3 NATURE OF THE ASSAY -- 24.4 APPLICATIONS OF CYTOTOXICITY ASSAYS -- 24.5 GENOTOXICITY -- 24.6 INFLAMMATION -- REFERENCES -- CHAPTER 25 Culture of Specific Cell Types -- 25.1 CELL CULTURE OF SPECIALIZED CELLS -- 25.2 EPITHELIAL CELLS -- 25.3 MESENCHYMAL CELLS -- 25.4 NEUROECTODERMAL CELLS -- 25.5 HEMATOPOIETIC CELLS -- 25.6 PRODUCTION OF MONOCLONAL ANTIBODIES -- 25.7 GONADS -- 25.8 CULTURE OF CELLS FROM POIKILOTHERMS -- 25.9 TUMOR CELL CULTURE -- 25.10 SAMPLING -- 25.11 DISAGGREGATION -- 25.12 PRIMARY CULTURE -- 25.13 SELECTIVE CULTURE OF TUMOR CELLS -- 25.14 DEVELOPMENT OF CELL LINES -- 25.15 CHARACTERIZATION OF TUMOR CELL CULTURES -- 25.16 SPECIFIC TUMOR TYPES -- REFERENCES -- CHAPTER 26 Stem Cells -- 26.1 EMBRYONIC STEM CELLS -- 26.2 GERM CELLS -- 26.3 EXTRAEMBRYONIC CELLS -- 26.4 HEMOPOIETIC STEM CELLS -- 26.5 APPICATION OF STEM CELLS TO REGENERATIVE MEDICINE -- 26.6 CANCER STEM CELLS -- REFERENCES -- CHAPTER 27 Training Programs -- 27.1 OBJECTIVES -- 27.2 PREPARATIVE AND MANIPULATIVE SKILLS -- 27.3 BASIC CELL CULTURE TECHNIQUES -- 27.4 ADVANCED EXERCISES -- 27.5 SPECIALIZED EXERCISES -- REFERENCES -- CHAPTER 28 Problem Solving -- 28.1 ABNORMAL APPEARANCE OF CELLS -- 28.2 SLOW CELL GROWTH -- 28.3 MEDIUM -- 28.4 SUBSTRATES AND CONTAINERS -- 28.5 MICROBIAL CONTAMINATION -- 28.6 CHEMICAL CONTAMINATION -- 28.7 PRIMARY CULTURE -- 28.8 DIFFERENTIATION -- 28.9 FEEDING -- 28.10 SUBCULTURE -- 28.11 CLONING -- 28.12 CROSS-CONTAMINATION AND MISIDENTIFICATION -- 28.13 CRYOPRESERVATION -- 28.14 CELL COUNTING -- REFERENCE -- CHAPTER 29 In Conclusion. 327 $aAppendix I Calculations and preparation of reagents -- REFERENCES -- APPENDIX II Glossary -- REFERENCE -- Index -- Links -- 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