| Contents Notes |
Foreword -- Acknowledgments -- Abbreviations -- Book navigation -- Part I. Understanding Cell Culture. Chapter 1. Introduction. 1.1. Terminology -- 1.2. Historical development -- 1.3. Applications -- 1.4. Advantages of tissue culture -- 1.5. Limitations of tissue culture -- References -- Chapter 2. Biology of Cultured Cells. 2.1. The culture environment -- 2.2. Cell adhesion -- 2.3. Cell division -- 2.4. Cell fate -- 2.5. Cell death -- References -- Chapter 3. Origin and Evolution of Cultured Cells. 3.1. Origin of cultured cells -- 3.2. Evolution of cell lines -- 3.3. Changes in genotype -- 3.4. Changes in phenotype -- 3.5. Senescence and immortalization -- 3.6. Transformation -- 3.7. Conclusions : Origin and evolution -- References -- Part II. Laboratory and Regulatory Requirements. Chapter 4. Laboratory Design and Layout. 4.1. Design requirements -- 4.2. Layout of laboratory areas -- 4.3. Disaster and contingency planning -- References -- Chapter 5. Equipment and Materials. 5.1. Sterile handling area equipment -- 5.2. Imaging and analysis equipment -- 5.3. Incubation equipment -- 5.4. Preparation and washup equipment -- 5.5. Cold storage equipment -- References -- Chapter 6. Safety and Bioethics. 6.1. Laboratory safety -- 6.2. Hazards in tissue culture laboratories -- 6.3. Biosafety -- 6.4. Bioethics -- References -- Chapter 7. Reproducibility and Good Cell Culture Practice. 7.1. Reproducibility -- 7.2. Good practice requirements -- 7.3. Cell line provenance -- 7.4. Validation testing -- 7.5. Quality Assurance (QA) -- 7.6. Replicate sampling -- References -- Part III. Medium and Substrate Requirements. Chapter 8. Culture Vessels and Substrates. 8.1. Attachment and growth requirements -- 8.2. Substrate materials -- 8.3. Substrate treatments -- 8.4. Feeder layers -- 8.5. Choice of culture vessel -- 8.6. Application-specific vessels -- References -- Chapter 9. Defined Media and Supplements. 9.1. Medium development -- 9.2. Physicochemical properties -- 9.3. Balanced salt solutions -- 9.4. Media formulations -- 9.5. Serum -- 9.6. Other media supplements -- 9.7. Choice of complete medium -- 9.8. Storage of medium and serum -- References -- Chapter 10. Serum-Free Media. 10.1. Rationale for serum-free medium -- 10.2. Development of serum-free medium -- 10.3. Serum-free media formulations -- 10.4. Serum-free supplements -- 10.5. Serum replacements -- 10.6. Use of serum-free medium -- 10.7. Xeno-free media -- 10.8. Animal product-free media -- 10.9. Conclusions : Serum-free media -- References -- Chapter 11. Preparation and Sterilization. 11.1. Terminology : Preparation -- 11.2. Sterilization methods -- 11.3. Glassware -- 11.4. Other laboratory apparatus -- 11.5. Water -- 11.6. Media and other reagents -- 11.7. Sterile filtration -- 11.8. Medium testing -- References -- Part IV Handling Cultures. Chapter 12. Aseptic Technique. 12.1. Objectives of aseptic technique -- 12.2. Elements of aseptic environment -- 12.3. Sterile handling -- 12.4. Good aseptic technique -- 12.5. Controlling equipment contamination -- References -- Chapter 13. Primary Culture. 13.1. Rational for primary culture -- 13.2. Initiation of primary culture -- 13.3. Tissue acquisition and isolation -- 13.4. Primary explantation -- 13.5. Enzymatic disaggregation -- 13.6. Mechanical disaggregation -- 13.7. Enrichment of viable cells -- 13.8. Record keeping for primary culture -- 13.9. Conclusions : Primary culture -- References -- Chapter 14. Subculture and Cell Lines. 14.1. Terminology : Cell line and subculture -- 14.2. Initiating a cell line -- 14.3. Choosing a cell line -- 14.4. Maintaining a cell line -- 14.5. Replacing medium (Feeding )-- 14.6. Subculture (Passaging) -- 14.7. Maintaining suspension cultures -- 14.8. Serum-free subculture -- 14.9. Record keeping for cell lines -- References -- Chapter 15. Cryopreservation and Banking. 15.1. Principles of cryopreservation -- 15.2. Apparatus for cryopreservation -- 15.3. Requirements for cryopreservation -- 15.4. Cryopreservation procedures -- 15.5. Cell banking procedures -- 15.6. Cell repositories -- 15.7. Record keeping for frozen stocks -- 15.8. Transporting cells -- References -- Part V. Validation and Characterization. Chapter 16. Microbial Contamination. 16.1. Sources of contamination -- 16.2. Management of contamination -- 16.3. Visible microbial contamination -- 16.4. Mycoplasma contamination -- 16.5. Viral contamination -- 16.6. Dealing with persistent contamination -- References -- Chapter 17. Cell Line Misidentification and Authentication. 17.1. Terminology : Cross-contamination, misidentification, and authentication -- 17.2. Misidentified cell lines -- 17.3. Cell line authentication -- 17.4. Authentication of challenging samples -- 17.5. Conclusions : Authentication -- References -- Chapter 18. Cell Line Characterization. 18.1. Priorities and essential characterization -- 18.2. Genotype-based characterization -- 18.3. Phenoype-based characterization -- 18.4. Cell imaging -- 18.5. Cell staining -- References -- Chapter 19. Quantitation and Growth Kinetics. 19.1. Cell counting -- 19.2. Cell viability -- 19.3. Cell proliferation -- 19.4. Cloning efficiency -- 19.5. DNA synthesis -- 19.6. Cell cycle analysis -- References -- Part VI. Physical and Genetic Manipulation. Chapter 20. Cell Cloning and Selection. 20.1. Terminology : Cloning and selection -- 20.2. Cloning by limiting dilution -- 20.3. Cloning in suspension -- 20.4. Selection of clones -- 20.5. Replica plating -- 20.6. Stimulation of cloning efficiency -- 20.7. Selective culture conditions -- 20.8. Conclusions : Cloning and selection -- References -- Chapter 21. Cell Separation and Sorting. 21.1. Cell density and isopycnic centrifugation -- 21.2. Cell size and sedimentation velocity -- 21.3. Magnetic separation and sorting -- 21.4. Fluorescence-Activated Cell Sorting (FACS) -- 21.5. Microfluidic sorting -- 21.6. Conclusions : Sorting and separation -- References -- Chapter 22. Genetic Modification and Immortalization. 22.1. Gene delivery -- 22.2. Gene editing -- 22.3. Immortalization -- 22.4. Screening and artifacts -- References -- Part VII. Stem Cells and Differentiated Cells. Chapter 23. Culture of Stem Cells. 23.1. Terminology : Stem cells -- 23.2. Embryonic Stem Cells (ESCs) -- 23.3. Induction of pluripotency -- 23.4. Human Pluripotent Stem Cell (hPSC) lines -- 23.5. Perinatal stem cells -- 23.6. Adult stem cells -- 23.7. Stem cell characterization and banking -- 23.8. Conclusions : Culture of stem cells -- References -- Chapter 24. Culture of Specific Cell Types. 24.1. Specialized cells and their availability -- 24.2. Epithelial cells -- 24.3. Mesenchymal cells -- 24.4. Neuroectodermal cells -- 24.5. Hematopoietic cells -- 24.6. Culture of cells from poikilotherms -- References -- Chapter 25. Culture of Tumor Cells. 25.1. Challenges of tumor cell culture -- 25.2. Primary culture of tumor cells -- 25.3. Development of tumor cell lines -- 25.4. Selective culture of tumor cells -- 25.5. Specific tumor types -- 25.6. Cancer Stem Cells (CSCs) -- References -- Chapter 26. Differentiation. 26.1. In Vitro models of differentiation -- 26.2. Differentiation status in culture -- 26.3. Induction of differentiation -- 26.4. Practical aspects -- 26.5. Ongoing challenges -- References -- Part VIII. Model Environments and Applications. Chapter 27. Three-Dimensional Culture. 27.1. Terminology : 3D culture -- 27.2. Technologies for 3D culture -- 27.3. Benefits and limitations of 3D culture -- 27.4. Scaffold-free 3D culture systems -- 27.5. Scaffold-based 3D culture systems -- 27.6. Organoid culture -- 27.7. Organotypic culture -- 27.8. Organ culture -- 27.9. Characterization of 3D cultures -- References -- Chapter 28. Scale-Up and Automation. 28.1. Terminology : Scale-up and bioreactors -- 28.2. Scale-up in suspension -- 28.3. Scale-up in monolayer -- 28.4. Monitoring and process control -- 28.5. Scale-up for manufacture -- 28.6. High-throughput screening -- 28.7. Automation and bioprinting -- References -- Chapter 29. Toxicity Testing. 29.1. In Vitro toxicity testing -- 29.2. Cytotoxicity assays -- 29.3. Genotoxicity assays -- 29.4. Carcinogenicity assays -- 29.5. Advanced models for toxicity testing -- References -- Part IX. Teaching and Troubleshooting. Chapter 30. Training. 30.1. Training principles -- 30.2. Training programs -- References -- Chapter 31. Problem Solving. 31.1. Microbial contamination -- 31.2. Cross-contamination and misidentification -- 31.3. Chemical contamination -- 31.4. Slow cell growth -- 31.5. Abnormal cell appearance -- 31.6. Problems with materials -- 31.7. Problems with primary culture -- 31.8. Problems with feeding or subculture -- 31.9. Problems with cryopreservation -- 31.10. Problems with cloning -- References -- Chapter 32. In Conclusion. Appendix A. Glossary -- Appendix B. Calculations and preparation of reagents. Calculations -- Preparation of reagents -- References -- Appendix C. Media formulations -- References -- Index. "Cell culture is the process used to keep cells alive in the laboratory. The process has undergone many changes since its discovery in 1907, as better techniques, equipment, and reagents continue to be developed. Cell culture now relies on a toolkit of techniques (e.g. aseptic technique), equipment (e.g. biosafety cabinet), and reagents (e.g. flasks and growth medium). Cells can be maintained and grown from many species and tissues, ranging from humans to fish, and from skin to brain. New methods and uses are emerging, allowing scientists to grow stem cells and develop more effective models through three-dimensional culture. However, cell culture can be challenging. Problems can affect any cell culture practitioner, even the most seasoned expert. Training is needed to ensure aseptic conditions, skilled handling, and an awareness of safety issues and of the purity and identity of the material being handled"-- "The 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. 'Freshney's Culture of Animal Cells' 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. The 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 : provides comprehensive single-volume coverage of basic skills and protocols, specialized techniques and applications, and new and emerging developments in the field ; covers 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 ; features a wealth of new content including protocols for gene delivery, iPSC generation and culture, and tumor spheroid formation. The eight edition of 'Freshney's Culture of Animal Cells' 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."--taken from back cover. |
