Biology

Freshney’s Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications, 8th Edition

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By Amanda Capes-Davis and R. Ian Freshney

Freshney’s Culture of Animal Cells PDF 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
Includes an updated and expanded companion website containing figures, artwork, and supplementary protocols to download and print

The eighth 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.

Table of Contents

Part I: Understanding Cell Culture

Introduction
- Terminology
- Historical Development
- Applications
- Advantages of Tissue Culture
- Limitations of Tissue Culture
- References
Biology of Cultured Cells
- The Culture Environment
- Cell Adhesion
- Cell Division
- Cell Fate
- Cell Death
- References
Origin and Evolution of Cultured Cells
- Origin of Cultured Cells
- Evolution of Cell Lines
- Changes in Genotype and Phenotype
- Senescence and Immortalization
- Transformation
- Conclusions
- References

Part II: Laboratory and Regulatory Requirements

Laboratory Design and Layout
- Design and Layout Requirements
- Disaster and Contingency Planning
- References
Equipment and Materials
- Sterile Handling Area
- Imaging, Incubation, and Storage Equipment
- References
Safety and Bioethics
- Laboratory Safety
- Biosafety and Bioethics
- References
Reproducibility and Good Cell Culture Practice
- Reproducibility Requirements
- Good Practice and Quality Assurance
- References

Part III: Medium and Substrate Requirements

Culture Vessels and Substrates
- Growth Requirements and Treatments
- Choice of Culture Vessel
- References
Defined Media and Supplements
- Development and Properties
- Media Formulations and Supplements
- References
Serum-Free Media
- Rationale and Development
- Formulations and Replacements
- References
Preparation and Sterilization
- Sterilization Methods and Protocols
- References

Part IV: Handling Cultures

Aseptic Technique
- Objectives and Best Practices
- Equipment Contamination Control
- References
Primary Culture
- Initiation and Explantation Techniques
- Enzymatic and Mechanical Disaggregation
- Record Keeping
- References
Subculture and Cell Lines
- Maintaining and Subculturing Cell Lines
- Record Keeping
- References
Cryopreservation and Banking
- Principles and Procedures
- Transporting Cells
- References

Part V: Validation and Characterization

Microbial Contamination
- Sources and Management
- Protocols for Disposal
- References
Cell Line Misidentification and Authentication
- Cross-Contamination and Provenance
- References
Cell Line Characterization
- Genotype and Phenotype-Based Methods
- References
Quantitation and Growth Kinetics
- Cell Counting and Proliferation Analysis
- References

Part VI: Physical and Genetic Manipulation

Cell Cloning and Selection
- Cloning Techniques and Selective Culture
- References
Cell Separation and Sorting
- Methods Including FACS and Magnetic Sorting
- References
Genetic Modification and Immortalization
- Gene Delivery and Editing Techniques
- References

Part VII: Stem Cells and Differentiated Cells

Culture of Stem Cells
- Types and Protocols
- References
Culture of Specific Cell Types
- Specialized Cells and Availability
- References
Culture of Tumor Cells
- Tumor Cell Line Development
- References
Differentiation
- Models and Practical Aspects
- References

Part VIII: Model Environments and Applications

Three-Dimensional Culture
- Technologies and Applications
- References
Scale-Up and Automation
- Bioreactors and High-Throughput Systems
- References
Toxicity Testing
- Cytotoxicity and Genotoxicity Assays
- References