Freshney's culture of animal cells : a manual of basic technique and specialized applications
Amanda Capes-Davis (Author, Editor), R. Ian Freshney (Author, Editor), Robert J. Geraghty (Editor), Raymond W. Nims (Editor)
"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"-- Provided by publisher
Print Book, English, 2021
Eighth edition View all formats and editions
Wiley Blackwell, Hoboken, NJ, 2021
Laboratory Manual
xxix, 796 pages : illustrations (chiefly color) ; 29 cm
9781119513018, 1119513014
1163959689
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
Revised edition of: Culture of animal cells : a manual of basic technique and specialized applications by R. Ian Freshney. Seventh edition. [2016]