Culture of animal cells (6th Ed.)
Auteurs : FRESHNEY, FRESHNEY
Since publication of the previous edition of this benchmark text, numerous types of equipment and materials have changed and new specialized techniques have been developed, particularly in the area of stem cell research and tissue engineering. These and other developments have been incorporated into this rigorously revised and expanded sixth edition of Culture of Animal Cells, the leading text in the basic methodology of cell culture. This authoritative volume offers a complete background related to growth of animal cells in culture. It contains many specialized protocols and advice on how to apply some of the basic protocols to teaching cell culture. Beginning with laboratory design, safety, validation, and bioethics, then continuing with preparation of media, primary culture, and cell lines through to characterization and authentication, contamination, specialized techniques, and troubleshooting, the unparalleled coverage includes: An all-new section addressing the additional caution required once a primary culture is converted to a cell line. Differentiation in stem cells and reprogramming of adult cells to form stem cells. A new section and protocol on cryopreservation by vitrification. Separate sections dealing with epithelial cells, mesenchymal cells, neurectodermal cells, and hematopoietic cells, a new chapter on germs cells/stemcells/amniocytes. A new section on organotypic culture. Increased emphasis on contamination and misidentification (including cross-contamination) with revised protocols for detection and elimination. A problem-solving chapter, glossary of terms, advice on sourcing materials, and list of suppliers. This landmark text presents the most accessible and comprehensive introduction available to the culture and experimental manipulation of animal cells and is indispensable for those in the field, including clinical and biopharmaceutical researchers and scientists, graduate students, laboratory managers in cell and molecular biology labs and genetics labs, trainees, and technicians.
List of Color Plates.
List of Protocols.
Preface and Acknowledgements.
Abbreviations.
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.
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.
3 Laboratory Design, Layout, and Equipment.
3.1 Layout, Furnishing, and Services.
3.2 Layout.
4 Equipment and Materials.
4.1 Requirements of a Tissue Culture Laboratory.
4.2 Aseptic Area.
4.3 Incubation and Culture.
4.4 Preparation and Sterilization.
4.5 Storage.
4.6 Supplementary Laboratory Equipment.
4.7 Specialized Equipment.
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.
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.
6.11 Validation.
7 Culture Vessels and Substrates.
7.1 The Substrate.
7.2 Treated Surfaces.
7.3 Choice of Culture Vessel.
7.4 Specialized Systems.
8 Defined Media and Supplements.
8.1 Development of Media.
8.2 Physicochemical Properties.
8.3 Balanced Salt Solutions.
8.4 Complete Media.
8.5 Serum.
8.6 Selection of Medium and Serum.
8.7 Other Supplements.
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 Selection of Serum-Free Medium.
9.6 Development of Serum-Free Medium.
9.7 Preparation of Serum-Free Medium.
9.8 Animal Protein-Free Media.
9.9 Conclusions.
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.
11 Primary Culture.
11.1 Initiation of a Primary Cell Culture.
11.2 Isolation of the Tissue.
11.3 Types of Primary Culture.
12 Subculture and Cell Lines.
12.1 Subculture and Propagation.
12.2 Choosing a Cell Line.
12.3 Routine Maintenance.
12.4 Subculture.
13 Cloning and Selection.
13.1 Cell Cloning.
13.2 Stimulation of Plating Efficiency.
13.3 Suspension Cloning.
13.4 Isolation of Clones.
13.5 Replica Plating.
13.6 Selective Inhibitors.
13.7 Isolation of Genetic Variants.
13.8 Interaction with Substrate.
14 Cell Separation.
14.1 Cell Density and Isopyknic Sedimentation.
14.2 Cell Size and Sedimentation Velocity.
14.3...
Date de parution : 01-1970
Ouvrage de 732 p.
