{"product_id":"freshneys-culture-of-animal-cells-isbn-9781119513018","title":"Freshney's Culture of Animal Cells","description":"\u003cb\u003eFRESHNEY’S CULTURE OF ANIMAL CELLS\u003c\/b\u003e \u003cp\u003e\u003cb\u003eTHE NEW EDITION OF THE LEADING TEXT ON THE BASIC METHODOLOGY OF CELL CULTURE, FULLY UPDATED TO REFLECT NEW APPLICATIONS INCLUDING IPSCS, CRISPR, AND ORGAN-ON-CHIP TECHNOLOGIES\u003c\/b\u003e  \u003c\/p\u003e\u003cp\u003e\u003ci\u003eFreshney’s Culture of Animal Cells\u003c\/i\u003e is the most comprehensive and up-to-date resource on the principles, techniques, equipment, and applications in the field of cell and tissue culture. Explaining both how to do tissue culture and why a technique is done in a particular way, this classic text covers the biology of cultured cells, how to select media and substrates, regulatory requirements, laboratory protocols, aseptic technique, experimental manipulation of animal cells, and much more. \u003c\/p\u003e\u003cp\u003eThe eighth edition contains extensively revised material that reflects the latest techniques and emerging applications in cell culture, such as the use of CRISPR\/Cas9 for gene editing and the adoption of chemically defined conditions for stem cell culture. A brand-new chapter examines the origin and evolution of cell lines, joined by a dedicated chapter on irreproducible research, its causes, and the importance of reproducibility and good cell culture practice. Throughout the book, updated chapters and protocols cover topics including live-cell imaging, 3D culture, scale-up and automation, microfluidics, high-throughput screening, and toxicity testing. This landmark text: \u003c\/p\u003e\u003cli\u003e\u003cbl\u003eProvides comprehensive single-volume coverage of basic skills and protocols, specialized techniques and applications, and new and emerging developments in the field\u003c\/bl\u003e\u003c\/li\u003e \u003cli\u003e\u003cbl\u003eCovers every essential area of animal cell culture, including lab design, disaster and contingency planning, safety, bioethics, media preparation, primary culture, mycoplasma and authentication testing, cell line characterization and cryopreservation, training, and troubleshooting\u003c\/bl\u003e\u003c\/li\u003e \u003cli\u003e\u003cbl\u003eFeatures a wealth of new content including protocols for gene delivery, iPSC generation and culture, and tumor spheroid formation\u003c\/bl\u003e\u003c\/li\u003e \u003cli\u003e\u003cbl\u003eIncludes an updated and expanded companion website containing figures, artwork, and supplementary protocols to download and print\u003c\/bl\u003e\u003c\/li\u003e \u003cp\u003eThe eighth edition of \u003ci\u003eFreshney’s Culture of Animal Cells\u003c\/i\u003e is an indispensable volume for anyone involved in the field, including undergraduate and graduate students, clinical and biopharmaceutical researchers, bioengineers, academic research scientists, and managers, technicians, and trainees working in cell biology, molecular biology, and genetics laboratories. \u003c\/p\u003e\u003cp\u003eForeword xix\u003c\/p\u003e \u003cp\u003eAcknowledgments xxi\u003c\/p\u003e \u003cp\u003eAbbreviations xxiii\u003c\/p\u003e \u003cp\u003eBook Navigation xxix\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart I Understanding Cell Culture 1\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e1. Introduction 3\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e1.1 Terminology 3\u003c\/p\u003e \u003cp\u003e1.2 Historical Development 4\u003c\/p\u003e \u003cp\u003e1.3 Applications 12\u003c\/p\u003e \u003cp\u003e1.4 Advantages of Tissue Culture 13\u003c\/p\u003e \u003cp\u003e1.5 Limitations of Tissue Culture 15\u003c\/p\u003e \u003cp\u003eReferences 18\u003c\/p\u003e \u003cp\u003e\u003cb\u003e2. Biology of Cultured Cells 23\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e2.1 The Culture Environment 23\u003c\/p\u003e \u003cp\u003e2.2 Cell Adhesion 23\u003c\/p\u003e \u003cp\u003e2.3 Cell Division 28\u003c\/p\u003e \u003cp\u003e2.4 Cell Fate 30\u003c\/p\u003e \u003cp\u003e2.5 Cell Death 35\u003c\/p\u003e \u003cp\u003eReferences 36\u003c\/p\u003e \u003cp\u003e\u003cb\u003e3. Origin and Evolution of Cultured Cells 39\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e3.1 Origin of Cultured Cells 39\u003c\/p\u003e \u003cp\u003e3.2 Evolution of Cell Lines 40\u003c\/p\u003e \u003cp\u003e3.3 Changes in Genotype 43\u003c\/p\u003e \u003cp\u003e3.4 Changes in Phenotype 46\u003c\/p\u003e \u003cp\u003e3.5 Senescence and Immortalization 48\u003c\/p\u003e \u003cp\u003e\u003ci\u003eMinireview M3.1 Senescence and Immortalization 48\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e3.6 Transformation 50\u003c\/p\u003e \u003cp\u003e3.7 Conclusions: Origin and Evolution 58\u003c\/p\u003e \u003cp\u003eReferences 58\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart II Laboratory and Regulatory Requirements 63\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e4. Laboratory Design and Layout 65\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e4.1 Design Requirements 65\u003c\/p\u003e \u003cp\u003e4.2 Layout of Laboratory Areas 74\u003c\/p\u003e \u003cp\u003e4.3 Disaster and Contingency Planning 80\u003c\/p\u003e \u003cp\u003eReferences 83\u003c\/p\u003e \u003cp\u003e\u003cb\u003e5. Equipment and Materials 85\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e5.1 Sterile Handling Area Equipment 85\u003c\/p\u003e \u003cp\u003e5.2 Imaging and Analysis Equipment 97\u003c\/p\u003e \u003cp\u003e5.3 Incubation Equipment 99\u003c\/p\u003e \u003cp\u003e5.4 Preparation and Washup Equipment 104\u003c\/p\u003e \u003cp\u003e5.5 Cold Storage Equipment 107\u003c\/p\u003e \u003cp\u003eReferences 109\u003c\/p\u003e \u003cp\u003e\u003cb\u003e6. Safety and Bioethics 111\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e6.1 Laboratory Safety 111\u003c\/p\u003e \u003cp\u003e6.2 Hazards in Tissue Culture Laboratories 117\u003c\/p\u003e \u003cp\u003e6.3 Biosafety 121\u003c\/p\u003e \u003cp\u003e6.4 Bioethics 129\u003c\/p\u003e \u003cp\u003eReferences 132\u003c\/p\u003e \u003cp\u003e\u003cb\u003e7. Reproducibility and Good Cell Culture Practice 137\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e7.1 Reproducibility 137\u003c\/p\u003e \u003cp\u003e7.2 Good Practice Requirements 141\u003c\/p\u003e \u003cp\u003e7.3 Cell Line Provenance 145\u003c\/p\u003e \u003cp\u003e7.4 Validation Testing 146\u003c\/p\u003e \u003cp\u003e7.5 Quality Assurance (QA) 148\u003c\/p\u003e \u003cp\u003e7.6 Replicate Sampling 150\u003c\/p\u003e \u003cp\u003eReferences 151\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart III Medium and Substrate Requirements 155\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e8. Culture Vessels and Substrates 157\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e8.1 Attachment and Growth Requirements 157\u003c\/p\u003e \u003cp\u003e8.2 Substrate Materials 158\u003c\/p\u003e \u003cp\u003e8.3 Substrate Treatments 159\u003c\/p\u003e \u003cp\u003e8.4 Feeder Layers 163\u003c\/p\u003e \u003cp\u003e8.5 Choice of Culture Vessel 164\u003c\/p\u003e \u003cp\u003e8.6 Application-Specific Vessels 170\u003c\/p\u003e \u003cp\u003eReferences 173\u003c\/p\u003e \u003cp\u003e\u003cb\u003e9. Defined Media and Supplements 177\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e9.1 Medium Development 177\u003c\/p\u003e \u003cp\u003e9.2 Physicochemical Properties 177\u003c\/p\u003e \u003cp\u003e9.3 Balanced Salt Solutions 185\u003c\/p\u003e \u003cp\u003e9.4 Media Formulations 186\u003c\/p\u003e \u003cp\u003e9.5 Serum 189\u003c\/p\u003e \u003cp\u003e9.6 Other Media Supplements 191\u003c\/p\u003e \u003cp\u003e9.7 Choice of Complete Medium 191\u003c\/p\u003e \u003cp\u003e9.8 Storage of Medium and Serum 194\u003c\/p\u003e \u003cp\u003eSuppliers 194\u003c\/p\u003e \u003cp\u003eReferences 194\u003c\/p\u003e \u003cp\u003e\u003cb\u003e10. Serum-Free Media 199\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e10.1 Rationale for Serum-Free Medium 199\u003c\/p\u003e \u003cp\u003e10.2 Development of Serum-Free Medium 201\u003c\/p\u003e \u003cp\u003e10.3 Serum-Free Media Formulations 202\u003c\/p\u003e \u003cp\u003e10.4 Serum-Free Supplements 203\u003c\/p\u003e \u003cp\u003e10.5 Serum Replacements 209\u003c\/p\u003e \u003cp\u003e10.6 Use of Serum-Free Medium 209\u003c\/p\u003e \u003cp\u003e10.7 Xeno-Free Media 213\u003c\/p\u003e \u003cp\u003e10.8 Animal Product-Free Media 214\u003c\/p\u003e \u003cp\u003e10.9 Conclusions: Serum-Free Media 214\u003c\/p\u003e \u003cp\u003eSuppliers 214\u003c\/p\u003e \u003cp\u003eReferences 215\u003c\/p\u003e \u003cp\u003e\u003cb\u003e11. Preparation and Sterilization 219\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e11.1 Terminology: Preparation 219\u003c\/p\u003e \u003cp\u003e11.2 Sterilization Methods 220\u003c\/p\u003e \u003cp\u003e11.3 Glassware 224\u003c\/p\u003e \u003cp\u003e\u003ci\u003eProtocol P11.1 Preparation and Sterilization of Glassware 224\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e11.4 Other Laboratory Apparatus 229\u003c\/p\u003e \u003cp\u003e11.5 Water 229\u003c\/p\u003e \u003cp\u003e11.6 Media and Other Reagents 233\u003c\/p\u003e \u003cp\u003e11.7 Sterile Filtration 238\u003c\/p\u003e \u003cp\u003e11.8 Medium Testing 242\u003c\/p\u003e \u003cp\u003eSuppliers 247\u003c\/p\u003e \u003cp\u003eReferences 247\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart IV Handling Cultures 249\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e12. Aseptic Technique 251\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e12.1 Objectives of Aseptic Technique 251\u003c\/p\u003e \u003cp\u003e12.2 Elements of Aseptic Environment 252\u003c\/p\u003e \u003cp\u003e12.3 Sterile Handling 258\u003c\/p\u003e \u003cp\u003e12.4 Good Aseptic Technique 260\u003c\/p\u003e \u003cp\u003e12.5 Controlling Equipment Contamination 265\u003c\/p\u003e \u003cp\u003eSuppliers 267\u003c\/p\u003e \u003cp\u003eReferences 267\u003c\/p\u003e \u003cp\u003e\u003cb\u003e13. Primary Culture 269\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e13.1 Rationale for Primary Culture 269\u003c\/p\u003e \u003cp\u003e13.2 Initiation of Primary Culture 270\u003c\/p\u003e \u003cp\u003e13.3 Tissue Acquisition and Isolation 274\u003c\/p\u003e \u003cp\u003e13.4 Primary Explantation 281\u003c\/p\u003e \u003cp\u003e\u003ci\u003eProtocol P13.3 Culture of Primary Explants 281\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e13.5 Enzymatic Disaggregation 283\u003c\/p\u003e \u003cp\u003e13.6 Mechanical Disaggregation 290\u003c\/p\u003e \u003cp\u003e\u003ci\u003eProtocol P13.7 Mechanical Disaggregation by Sieving 291\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e13.7 Enrichment of Viable Cells 292\u003c\/p\u003e \u003cp\u003e\u003ci\u003eProtocol P13.8 Enrichment of Viable Cells 292\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e13.8 Record Keeping for Primary Culture 293\u003c\/p\u003e \u003cp\u003e13.9 Conclusions: Primary Culture 294\u003c\/p\u003e \u003cp\u003eSuppliers 294\u003c\/p\u003e \u003cp\u003eReferences 294\u003c\/p\u003e \u003cp\u003e\u003cb\u003e14. Subculture and Cell Lines 297\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e14.1 Terminology: Cell Line and Subculture 297\u003c\/p\u003e \u003cp\u003e14.2 Initiating a Cell Line 298\u003c\/p\u003e \u003cp\u003e14.3 Choosing a Cell Line 300\u003c\/p\u003e \u003cp\u003e14.4 Maintaining a Cell Line 304\u003c\/p\u003e \u003cp\u003e14.5 Replacing Medium (Feeding) 309\u003c\/p\u003e \u003cp\u003e14.6 Subculture (Passaging) 312\u003c\/p\u003e \u003cp\u003e14.7 Maintaining Suspension Cultures 320\u003c\/p\u003e \u003cp\u003e14.8 Serum-Free Subculture 322\u003c\/p\u003e \u003cp\u003e14.9 Record Keeping for Cell Lines 323\u003c\/p\u003e \u003cp\u003eSuppliers 324\u003c\/p\u003e \u003cp\u003eReferences 325\u003c\/p\u003e \u003cp\u003e\u003cb\u003e15. Cryopreservation and Banking 327\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e15.1 Principles of Cryopreservation 327\u003c\/p\u003e \u003cp\u003e15.2 Apparatus for Cryopreservation 329\u003c\/p\u003e \u003cp\u003e15.3 Requirements for Cryopreservation 335\u003c\/p\u003e \u003cp\u003e15.4 Cryopreservation Procedures 336\u003c\/p\u003e \u003cp\u003e15.5 Cell Banking Procedures 341\u003c\/p\u003e \u003cp\u003e15.6 Cell Repositories 342\u003c\/p\u003e \u003cp\u003e15.7 Record Keeping for Frozen Stocks 345\u003c\/p\u003e \u003cp\u003e15.8 Transporting Cells 347\u003c\/p\u003e \u003cp\u003eSuppliers 348\u003c\/p\u003e \u003cp\u003eReferences 348\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart V Validation and Characterization 351\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e16. Microbial Contamination 353\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e16.1 Sources of Contamination 353\u003c\/p\u003e \u003cp\u003e16.2 Management of Contamination 359\u003c\/p\u003e \u003cp\u003e\u003ci\u003eProtocol P16.1 Disposal of Contaminated Cultures 360\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e16.3 Visible Microbial Contamination 361\u003c\/p\u003e \u003cp\u003e16.4 Mycoplasma Contamination 364\u003c\/p\u003e \u003cp\u003e16.5 Viral Contamination 373\u003c\/p\u003e \u003cp\u003e16.6 Dealing with Persistent Contamination 376\u003c\/p\u003e \u003cp\u003eSuppliers 376\u003c\/p\u003e \u003cp\u003eReferences 376\u003c\/p\u003e \u003cp\u003e\u003cb\u003e17. Cell Line Misidentification and Authentication 381\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e17.1 Terminology: Cross-Contamination, Misidentification, and Authentication 381\u003c\/p\u003e \u003cp\u003e17.2 Misidentified Cell Lines 382\u003c\/p\u003e \u003cp\u003e17.3 Cell Line Authentication 386\u003c\/p\u003e \u003cp\u003e17.4 Authentication of Challenging Samples 401\u003c\/p\u003e \u003cp\u003e17.5 Conclusions: Authentication 403\u003c\/p\u003e \u003cp\u003eSuppliers 403\u003c\/p\u003e \u003cp\u003eReferences 403\u003c\/p\u003e \u003cp\u003e\u003cb\u003e18. Cell Line Characterization 409\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e18.1 Priorities and Essential Characterization 409\u003c\/p\u003e \u003cp\u003e18.2 Genotype-Based Characterization 416\u003c\/p\u003e \u003cp\u003e18.3 Phenotype-Based Characterization 419\u003c\/p\u003e \u003cp\u003e18.4 Cell Imaging 423\u003c\/p\u003e \u003cp\u003e18.5 Cell Staining 428\u003c\/p\u003e \u003cp\u003eSuppliers 430\u003c\/p\u003e \u003cp\u003eReferences 430\u003c\/p\u003e \u003cp\u003e\u003cb\u003e19. Quantitation and Growth Kinetics 437\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e19.1 Cell Counting 437\u003c\/p\u003e \u003cp\u003e19.3 Cell Proliferation 450\u003c\/p\u003e \u003cp\u003e19.4 Cloning Efficiency 456\u003c\/p\u003e \u003cp\u003e19.5 DNA Synthesis 460\u003c\/p\u003e \u003cp\u003e19.6 Cell Cycle Analysis 461\u003c\/p\u003e \u003cp\u003eSuppliers 461\u003c\/p\u003e \u003cp\u003eReferences 461\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart VI Physical and Genetic Manipulation 465\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e20. Cell Cloning and Selection 467\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e20.1 Terminology: Cloning and Selection 467\u003c\/p\u003e \u003cp\u003e20.2 Cloning by Limiting Dilution 468\u003c\/p\u003e \u003cp\u003e20.3 Cloning in Suspension 473\u003c\/p\u003e \u003cp\u003e20.4 Selection of Clones 477\u003c\/p\u003e \u003cp\u003e20.5 Replica Plating 480\u003c\/p\u003e \u003cp\u003e20.6 Stimulation of Cloning Efficiency 481\u003c\/p\u003e \u003cp\u003e20.7 Selective Culture Conditions 485\u003c\/p\u003e \u003cp\u003e20.8 Conclusions: Cloning and Selection 487\u003c\/p\u003e \u003cp\u003eSuppliers 487\u003c\/p\u003e \u003cp\u003eReferences 487\u003c\/p\u003e \u003cp\u003e\u003cb\u003e21. Cell Separation and Sorting 491\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e21.1 Cell Density and Isopycnic Centrifugation 491\u003c\/p\u003e \u003cp\u003e21.2 Cell Size and Sedimentation Velocity 495\u003c\/p\u003e \u003cp\u003e21.3 Magnetic Separation and Sorting 496\u003c\/p\u003e \u003cp\u003e\u003ci\u003eProtocol P21.2 Magnet-Activated Cell Sorting (MACS) 499\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e21.4 Fluorescence-Activated Cell Sorting (FACS) 500\u003c\/p\u003e \u003cp\u003e21.5 Microfluidic Sorting 502\u003c\/p\u003e \u003cp\u003e\u003ci\u003eMinireview M21.1 Microfluidic Cell Culture 503\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e21.6 Conclusions: Sorting and Separation 505\u003c\/p\u003e \u003cp\u003eSuppliers 505\u003c\/p\u003e \u003cp\u003eReferences 505\u003c\/p\u003e \u003cp\u003e\u003cb\u003e22. Genetic Modification and Immortalization 509\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e22.1 Gene Delivery 509\u003c\/p\u003e \u003cp\u003e22.2 Gene Editing 517\u003c\/p\u003e \u003cp\u003e22.3 Immortalization 523\u003c\/p\u003e \u003cp\u003e22.4 Screening and Artifacts 526\u003c\/p\u003e \u003cp\u003eSuppliers 528\u003c\/p\u003e \u003cp\u003eReferences 528\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart VII Stem Cells and Differentiated Cells 535\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e23. Culture of Stem Cells 537\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e23.1 Terminology: Stem Cells 537\u003c\/p\u003e \u003cp\u003e23.2 Embryonic Stem Cells (ESCs) 540\u003c\/p\u003e \u003cp\u003e23.3 Induction of Pluripotency 545\u003c\/p\u003e \u003cp\u003e\u003ci\u003eProtocol P23.1 Generation of iPSCs Using Sendai Viral Vectors 547\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e23.4 Human Pluripotent Stem Cell (hPSC) Lines 549\u003c\/p\u003e \u003cp\u003e23.5 Perinatal Stem Cells 556\u003c\/p\u003e \u003cp\u003e23.6 Adult Stem Cells 557\u003c\/p\u003e \u003cp\u003e23.7 Stem Cell Characterization and Banking 558\u003c\/p\u003e \u003cp\u003e23.8 Conclusions: Culture of Stem Cells 560\u003c\/p\u003e \u003cp\u003eSuppliers 561\u003c\/p\u003e \u003cp\u003eReferences 561\u003c\/p\u003e \u003cp\u003e\u003cb\u003e24. Culture of Specific Cell Types 567\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e24.1 Specialized Cells and Their Availability 567\u003c\/p\u003e \u003cp\u003e24.2 Epithelial Cells 572\u003c\/p\u003e \u003cp\u003e24.3 Mesenchymal Cells 577\u003c\/p\u003e \u003cp\u003e24.4 Neuroectodermal Cells 580\u003c\/p\u003e \u003cp\u003e24.5 Hematopoietic Cells 581\u003c\/p\u003e \u003cp\u003e24.6 Culture of Cells from Poikilotherms 585\u003c\/p\u003e \u003cp\u003eSuppliers 587\u003c\/p\u003e \u003cp\u003eReferences 587\u003c\/p\u003e \u003cp\u003e\u003cb\u003e25. Culture of Tumor Cells 593\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e25.1 Challenges of Tumor Cell Culture 593\u003c\/p\u003e \u003cp\u003e25.2 Primary Culture of Tumor Cells 594\u003c\/p\u003e \u003cp\u003e25.3 Development of Tumor Cell Lines 596\u003c\/p\u003e \u003cp\u003e25.4 Selective Culture of Tumor Cells 599\u003c\/p\u003e \u003cp\u003e25.5 Specific Tumor Types 603\u003c\/p\u003e \u003cp\u003e25.6 Cancer Stem Cells (CSCs) 606\u003c\/p\u003e \u003cp\u003e\u003ci\u003eMinireview M25.1 Culture of Cancer Stem Cells 606\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003eSuppliers 608\u003c\/p\u003e \u003cp\u003eReferences 608\u003c\/p\u003e \u003cp\u003e\u003cb\u003e26. Differentiation 615\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e26.1 \u003ci\u003eIn Vitro \u003c\/i\u003eModels of Differentiation 615\u003c\/p\u003e \u003cp\u003e26.2 Differentiation Status in Culture 617\u003c\/p\u003e \u003cp\u003e26.3 Induction of Differentiation 620\u003c\/p\u003e \u003cp\u003e26.4 Practical Aspects 628\u003c\/p\u003e \u003cp\u003e26.5 Ongoing Challenges 629\u003c\/p\u003e \u003cp\u003eSuppliers 631\u003c\/p\u003e \u003cp\u003eReferences 631\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart VIII Model Environments and Applications 639\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e27. Three-Dimensional Culture 641\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e27.1 Terminology: 3D Culture 641\u003c\/p\u003e \u003cp\u003e27.2 Technologies for 3D Culture 643\u003c\/p\u003e \u003cp\u003e\u003ci\u003eMinireview M27.1 Advances in Technologies Enabling 3D Cell Culture and the Formation of Tissue-Like Architecture \u003c\/i\u003eIn Vitro\u003ci\u003e 643\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e27.3 Benefits and Limitations of 3D Culture 646\u003c\/p\u003e \u003cp\u003e27.4 Scaffold-Free 3D Culture Systems 647\u003c\/p\u003e \u003cp\u003e27.5 Scaffold-Based 3D Culture Systems 652\u003c\/p\u003e \u003cp\u003e27.6 Organoid Culture 659\u003c\/p\u003e \u003cp\u003e27.7 Organotypic Culture 660\u003c\/p\u003e \u003cp\u003e27.8 Organ Culture 662\u003c\/p\u003e \u003cp\u003e27.9 Characterization of 3D Cultures 662\u003c\/p\u003e \u003cp\u003eSuppliers 663\u003c\/p\u003e \u003cp\u003eReferences 663\u003c\/p\u003e \u003cp\u003e\u003cb\u003e28. Scale-Up and Automation 669\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e28.1 Terminology: Scale-Up and Bioreactors 669\u003c\/p\u003e \u003cp\u003e28.2 Scale-Up in Suspension 671\u003c\/p\u003e \u003cp\u003e28.3 Scale-Up in Monolayer 677\u003c\/p\u003e \u003cp\u003e28.4 Monitoring and Process Control 685\u003c\/p\u003e \u003cp\u003e28.5 Scale-Up for Manufacture 688\u003c\/p\u003e \u003cp\u003e\u003ci\u003eMinireview M28.1 Culture Scale-Up and Bioreactors 688\u003c\/i\u003e\u003c\/p\u003e \u003cp\u003e28.6 High-Throughput Screening 691\u003c\/p\u003e \u003cp\u003e28.7 Automation and Bioprinting 691\u003c\/p\u003e \u003cp\u003eSuppliers 696\u003c\/p\u003e \u003cp\u003eReferences 696\u003c\/p\u003e \u003cp\u003e\u003cb\u003e29. Toxicity Testing 701\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e29.1 \u003ci\u003eIn Vitro \u003c\/i\u003eToxicity Testing 701\u003c\/p\u003e \u003cp\u003e29.2 Cytotoxicity Assays 704\u003c\/p\u003e \u003cp\u003e29.3 Genotoxicity Assays 715\u003c\/p\u003e \u003cp\u003e29.4 Carcinogenicity Assays 716\u003c\/p\u003e \u003cp\u003e29.5 Advanced Models for Toxicity Testing 716\u003c\/p\u003e \u003cp\u003eSuppliers 719\u003c\/p\u003e \u003cp\u003eReferences 719\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart IX Teaching and Troubleshooting 725\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e30. Training 727\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e30.1 Training Principles 727\u003c\/p\u003e \u003cp\u003e30.2 Training Programs 729\u003c\/p\u003e \u003cp\u003eReferences 731\u003c\/p\u003e \u003cp\u003e\u003cb\u003e31. Problem Solving 733\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e31.1 Microbial Contamination 733\u003c\/p\u003e \u003cp\u003e31.2 Cross-Contamination and Misidentification 737\u003c\/p\u003e \u003cp\u003e31.3 Chemical Contamination 738\u003c\/p\u003e \u003cp\u003e31.4 Slow Cell Growth 738\u003c\/p\u003e \u003cp\u003e31.5 Abnormal Cell Appearance 740\u003c\/p\u003e \u003cp\u003e31.6 Problems with Materials 741\u003c\/p\u003e \u003cp\u003e31.7 Problems with Primary Culture 744\u003c\/p\u003e \u003cp\u003e31.8 Problems with Feeding or Subculture 746\u003c\/p\u003e \u003cp\u003e31.9 Problems with Cryopreservation 748\u003c\/p\u003e \u003cp\u003e31.10 Problems with Cloning 750\u003c\/p\u003e \u003cp\u003eReferences 752\u003c\/p\u003e \u003cp\u003e\u003cb\u003e32. In Conclusion 753\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003eAppendix A Glossary 755\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003eAppendix B Calculations and Preparation of Reagents 761\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003eCalculations 761\u003c\/p\u003e \u003cp\u003eCounting Cells with a Hemocytometer 761\u003c\/p\u003e \u003cp\u003eDilution of a Cell Suspension 761\u003c\/p\u003e \u003cp\u003ePopulation Doubling Level (PDL) 761\u003c\/p\u003e \u003cp\u003eMolarity 762\u003c\/p\u003e \u003cp\u003ePercentages and Dilutions 762\u003c\/p\u003e \u003cp\u003ePressure 762\u003c\/p\u003e \u003cp\u003eRotor Speed (rpm to g) 762\u003c\/p\u003e \u003cp\u003ePreparation of Reagents 762\u003c\/p\u003e \u003cp\u003eAcetic Acid: Methanol 762\u003c\/p\u003e \u003cp\u003eAgar (2.5%) 762\u003c\/p\u003e \u003cp\u003eAlcohol (70%) 762\u003c\/p\u003e \u003cp\u003eBacto\u003csup\u003e™\u003c\/sup\u003e Peptone (5%) 763\u003c\/p\u003e \u003cp\u003eBalanced Salt Solutions 763\u003c\/p\u003e \u003cp\u003eCarboxymethylcellulose (CMC; 4%) 763\u003c\/p\u003e \u003cp\u003eChick Embryo Extract 763\u003c\/p\u003e \u003cp\u003eCollagenase 763\u003c\/p\u003e \u003cp\u003eCollection Medium 763\u003c\/p\u003e \u003cp\u003eCrystal Violet (0.1%) 764\u003c\/p\u003e \u003cp\u003eDexamethasone (1 mg\/ml) 764\u003c\/p\u003e \u003cp\u003eDissection Balanced Salt Solution (DBSS) 764\u003c\/p\u003e \u003cp\u003eDulbecco’s Phosphate-Buffered Saline Without Ca\u003csup\u003e2\u003c\/sup\u003e\u003csup\u003e+\u003c\/sup\u003e and Mg\u003csup\u003e2\u003c\/sup\u003e\u003csup\u003e+\u003c\/sup\u003e (DPBS-A) 764\u003c\/p\u003e \u003cp\u003eEDTA (10 mM in DPBS-A) 764\u003c\/p\u003e \u003cp\u003eEGTA 764\u003c\/p\u003e \u003cp\u003eErythrosin B 764\u003c\/p\u003e \u003cp\u003eGelatin (1%) 765\u003c\/p\u003e \u003cp\u003eGiemsa Stain 765\u003c\/p\u003e \u003cp\u003eGlucose (20%) 765\u003c\/p\u003e \u003cp\u003eGlutamine 200 mM 765\u003c\/p\u003e \u003cp\u003eHanks’s Balanced Salt Solution (HBSS) 765\u003c\/p\u003e \u003cp\u003eHAT Medium 765\u003c\/p\u003e \u003cp\u003eHB Medium 765\u003c\/p\u003e \u003cp\u003eHEPES 765\u003c\/p\u003e \u003cp\u003eHoechst 33258 766\u003c\/p\u003e \u003cp\u003eMedia 766\u003c\/p\u003e \u003cp\u003e2-Mercaptoethanol (𝛽-Mercaptoethanol; 0.1 M) 766\u003c\/p\u003e \u003cp\u003eMethylcellulose (Methocel, 1.6%) 766\u003c\/p\u003e \u003cp\u003eMitomycin C (100 μg\/ml) 766\u003c\/p\u003e \u003cp\u003eMTT (50 mg\/ml) 766\u003c\/p\u003e \u003cp\u003eN2 Supplement 766\u003c\/p\u003e \u003cp\u003eN2B27 Medium 767\u003c\/p\u003e \u003cp\u003eNaphthalene Black (Amido Black; 1%) 767\u003c\/p\u003e \u003cp\u003eNon-essential Amino Acids (NEAA, 100×) 767\u003c\/p\u003e \u003cp\u003eParaformaldehyde (4%) 767\u003c\/p\u003e \u003cp\u003eTrypan Blue (0.4%) 767\u003c\/p\u003e \u003cp\u003eTrypsin (2.5%) 768\u003c\/p\u003e \u003cp\u003eVersene 768\u003c\/p\u003e \u003cp\u003eSuppliers 768\u003c\/p\u003e \u003cp\u003eReferences 768\u003c\/p\u003e \u003cp\u003e\u003cb\u003eAppendix C Media Formulations 769\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003eReferences 779\u003c\/p\u003e \u003cp\u003eIndex 781 \u003c\/p\u003e \u003cp\u003e\u003cb\u003eAMANDA CAPES-DAVIS, PHD,\u003c\/b\u003e is a cell culture scientist and technical writer. She was Founding Manager and Honorary Scientist at CellBank Australia, Children’s Medical Research Institute (CMRI), and is a member of the International Cell Line Authentication Committee (ICLAC). She was a Reviewing Editor for the 7th edition of \u003ci\u003eCulture of Animal Cells\u003c\/i\u003e, and has written numerous journal articles, policies, protocols, and white papers on good cell culture practice.\u003c\/p\u003e \u003cp\u003e\u003cb\u003eR. IAN FRESHNEY, PHD,\u003c\/b\u003e was an honorary Senior Research Fellow at the Institute of Cancer Sciences at the University of Glasgow, UK. Dr Freshney, who died in 2019, was a world-renowned cancer biologist and a pioneer in cell culture techniques who made important contributions to new approaches for treating cancer patients. He taught cell culture courses at national and international level, and wrote and edited numerous books, including the first seven editions of \u003ci\u003eCulture of Animal Cells\u003c\/i\u003e.  \u003c\/p\u003e\u003cp\u003e\u003cb\u003eTHE NEW EDITION OF THE LEADING TEXT ON THE BASIC METHODOLOGY OF CELL CULTURE, FULLY UPDATED TO REFLECT NEW APPLICATIONS INCLUDING IPSCS, CRISPR, AND ORGAN-ON-CHIP TECHNOLOGIES\u003c\/b\u003e  \u003c\/p\u003e\u003cp\u003e\u003ci\u003eFreshney’s Culture of Animal Cells\u003c\/i\u003e is the most comprehensive and up-to-date resource on the principles, techniques, equipment, and applications in the field of cell and tissue culture. Explaining both how to do tissue culture and why a technique is done in a particular way, this classic text covers the biology of cultured cells, how to select media and substrates, regulatory requirements, laboratory protocols, aseptic technique, experimental manipulation of animal cells, and much more. \u003c\/p\u003e\u003cp\u003eThe eighth edition contains extensively revised material that reflects the latest techniques and emerging applications in cell culture, such as the use of CRISPR\/Cas9 for gene editing and the adoption of chemically defined conditions for stem cell culture. A brand-new chapter examines the origin and evolution of cell lines, joined by a dedicated chapter on irreproducible research, its causes, and the importance of reproducibility and good cell culture practice. Throughout the book, updated chapters and protocols cover topics including live-cell imaging, 3D culture, scale-up and automation, microfluidics, high-throughput screening, and toxicity testing. This landmark text: \u003c\/p\u003e\u003cli\u003e\u003cbl\u003eProvides comprehensive single-volume coverage of basic skills and protocols, specialized techniques and applications, and new and emerging developments in the field\u003c\/bl\u003e\u003c\/li\u003e \u003cli\u003e\u003cbl\u003eCovers every essential area of animal cell culture, including lab design, disaster and contingency planning, safety, bioethics, media preparation, primary culture, mycoplasma and authentication testing, cell line characterization and cryopreservation, training, and troubleshooting\u003c\/bl\u003e\u003c\/li\u003e \u003cli\u003e\u003cbl\u003eFeatures a wealth of new content including protocols for gene delivery, iPSC generation and culture, and tumor spheroid formation\u003c\/bl\u003e\u003c\/li\u003e \u003cli\u003e\u003cbl\u003eIncludes an updated and expanded companion website containing figures, artwork, and supplementary protocols to download and print\u003c\/bl\u003e\u003c\/li\u003e \u003cp\u003eThe eighth edition of \u003ci\u003eFreshney’s Culture of Animal Cells\u003c\/i\u003e is an indispensable volume for anyone involved in the field, including undergraduate and graduate students, clinical and biopharmaceutical researchers, bioengineers, academic research scientists, and managers, technicians, and trainees working in cell biology, molecular biology, and genetics laboratories.\u003c\/p\u003e","brand":"Wiley-Blackwell","offers":[{"title":"Default Title","offer_id":47989245444325,"sku":"NP9781119513018","price":100.0,"currency_code":"USD","in_stock":false}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/1842\/7735\/files\/9781119513018.jpg?v=1761783359","url":"https:\/\/k12savings.com\/products\/freshneys-culture-of-animal-cells-isbn-9781119513018","provider":"K12savings","version":"1.0","type":"link"}