Editorial Reviews. About the Author. R. IAN FRESHNEY, PhD, is an honorary senior research Kindle Store; ›; Kindle eBooks; ›; Science & Math. Editorial Reviews. Review. "The book's sixth incarnation includes new sections on organotypic of Basic Technique and Specialized Applications - Kindle edition by R. Ian Freshney. Kindle Store; ›; Kindle eBooks; ›; Science & Math. Culture of Animal Cells. SIXTH EDITION. A Manual of Basic Technique and Specialized Applications. R. IAN FRESHNEY. #WILEY-BLACKWELL. WWW.
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Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications (6th ed.) by R. Ian Freshney. Read online. Read "Culture of Animal Cells A Manual of Basic Technique and Specialized Applications" by R. Ian Freshney available from Rakuten Kobo. Sign up today and. Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications, 7th Edition. Culture of Animal Cells: A Manual of Basic R. Ian Freshney.
Culture of Animal Cells: A Manual of Basic Technique. Ian Freshney. The fourth edition of Culture of Animal Cells: A Manual of Basic Technique offers the most complete training manual of its kind on the fundamental principles and techniques of animal cell culture. Within this volume, indispensable updates reflecting the latest progress in media, specialized techniques, biotechnology, DNA transfer, and tumor culture have been made. This edition has five new chapters expanding on serum-free media, scale-up and biofermentors, molecular techniques, immortalization, and troubleshooting.
Growth Cycle, Protocol Growth Curve with a Monolayer in Flasks, Protocol Growth Curve with a Monolayer in Multiwell Plates, Analysis of Monolayer Growth Curves, Suspension Cultures, Protocol Growth Curve with Cells in Suspension, Phases of the Growth Cycle, Derivatives from the Growth Curve, Analysis of Colony Formation, Automatic Colony Counting, Labeling Index, Protocol Labeling Index with [ 3H]Thymidine, Growth Fraction, Protocol Determination of Growth Fraction, Mitotic Index, Division Index, Cell Cycle Time, Cell Migration, Cytotoxicity, Viability, Toxicity, and Survival, In vitro Limitations, Pharmacokinetics, Metabolism, Tissue and Systemic Responses, Nature of the Assay, Viability, Protocol Estimation of Viability by Dye Exclusion, Protocol Estimation of Viability by Dye Uptake, Survival, Protocol Clonogenic Assay for Attached Cells, Assays Based on Cell Proliferation, Metabolic Cytotoxicity Assays, Microtitration Assays, Protocol Mtt-Based Cytotoxicity Assay, Comparison of Microtitration with Clonogenic Survival, Drug Interaction, Applications of Cytotoxicity Assays, Anticancer Drug Screening, Predictive Drug Testing for Tumors, Testing Pharmaceuticals, Genotoxicity, Sister Chromatid Exchange, Carcinogenicity, SpecializedCells, Cell Culture of Specialized Cells, Epithelial Cells, Epidermis, Protocol Epidermal Keratinocytes, Cornea, Protocol Corneal Epithelial Cells, Breast, Protocol Cervix, Protocol Cervical Epithelium, Gastrointestinal Tract, Protocol Isolation and Culture of Colonic Crypts, Liver, Hepatocyte Primary Cultures, Protocol Isolation of Rat Hepatocytes, Pancreas, Protocol Pancreatic Epithelium, Kidney, Protocol Kidney Epithelium, Bronchial and Tracheal Epithelium, Protocol Bronchial and Tracheal Epithelium, Oral Epithelium, Protocol Oral Keratinocytes, Prostate, Protocol Prostatic Epithelium, Mesenchymal Cells, Connective Tissue, Adipose Tissue, Protocol Primary Culture of Adipose Cells, Muscle, Protocol Cartilage, Protocol Chondrocytes in Alginate Beads, Bone, Protocol Osteoblasts, Endothelium, Protocol Isolation and Culture of Vascular Endothelial Cells, Neuroectodermal Cells, Neurons, Protocol Cerebellar Granule Cells, Glial Cells, Protocol Primary Culture of Human Astrocytes, Protocol Olfactory Ensheathing Cells, Endocrine Cells, Melanocytes, Protocol Culture of Melanocytes, Hematopoietic Cells, Gonads, Ovary, Testis, StemCells,GermCells,andAmniocytes, Stem Cells, Embryonic Stem Cells, Derivation of Human Embryonic Stem Cells, Passaging hES Cells, Protocol Manual Passage of hES Cells, Germ Cells, Extraembryonic Cells, Culture of Amniocytes, Protocol Culture of Amniocytes, Cells from Neonates and Juveniles, Multipotent Stem Cells from the Adult, Induced Pluripotent Stem Cells, Protocol Hematopoietic Colony-Forming Assays, Protocol Hematopoietic Colony-Forming Assays, CultureofTumorCells, Problems of Tumor Cell Culture, Sampling, Selection of Representative Cells, Preservation of Tissue by Freezing, Protocol Freezing Biopsies, Disaggregation, Primary Culture, Selective Culture of Tumor Cells, Selective Media, Suspension Cloning, Xenografts, Development of Cell Lines, Subculture of Primary Tumor Cultures, Continuous Cell Lines, Characterization of Tumor Cell Cultures, Heterogeneity of Tumor Cultures, Histotypic Culture, Culture of Mammary Tumor Cells, Lung, Stomach, Colon, Protocol Culture of Colorectal Tumors, Pancreas, Prostate, Bladder, Skin, Cervix, Glioma, Neuroblastoma, Seminoma, Lymphoma and Leukemia, Protocol Three-DimensionalCulture, Cell Interaction and Phenotypic Expression, Effect of Cell Density, Reciprocal Interactions, Choice of Models, Organ Culture, Gas and Nutrient Exchange, Structural Integrity, Growth and Differentiation, Limitations of Organ Culture, Types of Organ Culture, Protocol Gel and Sponge Techniques, Hollow Fibers, Spheroids, Protocol Rotating Chamber Systems, Immobilization of Living Cells in Alginate, Filter Well Inserts, Protocol Filter Well Inserts, Cultures of Neuronal Aggregates, Protocol Neuronal Aggregates, Organotypic Culture, Tissue Equivalents, Tissue Engineering, Imaging Cells in 3-D Constructs, Scale-upandAutomation, Scale-up in Suspension, Protocol Stirred 4-Liter Batch Suspension Culture, Continuous Culture, Scale and Complexity, Mixing and Aeration, Scale-up in Monolayer, Multisurface Propagators, Protocol Roller Culture, Protocol Roller Bottle Culture, Microcarriers, Protocol Microcarriers, Large Microcarriers, Perfused Monolayer Culture, Process Control, Automation, Robotic Cell Culture, High-Throughput Screening, SpecializedTechniques, Lymphocyte Preparation, Isolation by Density, Protocol Preparation of Lymphocytes, Blast Transformation, Protocol PHA Stimulation of Lymphocytes, Autoradiography, Protocol Microautoradiography, Time-Lapse Recording, Protocol Time-Lapse Video Recording, Cell Synchrony, Cell Separation, Blockade, Culture of Cells from Poikilotherms, Fish Cells, Insect Cells, Protocol Propagation of Insect Cells, Somatic Cell Fusion, Cell Hybridization, Protocol Cell Hybridization, Nuclear Transfer, Production of Monoclonal Antibodies, Protocol Production of Monoclonal Antibodies, TrainingPrograms, Objectives, Specialized Exercises, ProblemSolving, Abnormal Appearance of Cells, Slow Cell Growth, Problems Restricted to Your Own Stock, Medium, Formulation, Preparation, and Storage, Unstable Reagents, Purity of Medium Constituents, Substrates and Containers, Microbial Contamination, Widespread, Air Supply and Laminar-Flow Hoods, Chemical Contamination, Glassware, Pipettes, Cryopreservatives, Powders and Aerosols, Poor Take in Primary Culture, Wrong Cells Selected, Feeding, Regular Monolayers, Cell Cloning, Poor Take or Slow Growth, Uneven Growth, Cloning, Too Few Colonies per Dish, Too Many Colonies per Dish, Nonrandom Distribution, Poor Recovery, Changed Appearance after Cryopreservation, Loss of Stock, Hemocytometer, InConclusion, Appendix I: Glossary, Appendix V: ListofFigures 1.
Growth of Tissue Culture 1. Tissue Culture Applications 1.
Types of Tissue Culture 2. Cell Adhesion 2. Intercellular Junctions 2. A Cells Growing on Matrigel 2. Cell Cycle 2. Differentiation and Proliferation 2. Differentiation from Stem Cells 2. Commitment and Reversibility 2. Cell Interaction and Signaling 2. Evolution of a Cell Line 2.
Small Tissue Culture Laboratory 3. Medium-Sized Tissue Culture Laboratory 3. Large Tissue Culture Laboratory 3. Air Pressure Balance 3. Hot Room 3. Washingup Sink and Pipette Washer 3. Liquid-Nitrogen Store and Cryostore 4.
Laminar-Flow Hood 4. Pipette Controller 4. Pipettors 4. Graduated Bottle Dispenser 4. Syringe Dispensers 4. Automatic Dispensers 4. Plate Filler and Plate Reader 4.
Transfer Device 4. Aspiration of Medium 4. Inverted Microscope 4. Culture Chambers 4. CO2 Incubator 4. CO2 Incubator Design 4. Glassware Washing Machine 4. Bench-Top Autoclave 4. Freestanding Autoclave 4. Tubes 5. Probability of Contamination 5. Tissue Culture Work Area 5. Layout of Work Area 5. Layout of Horizontal Laminar-Flow Hood 5. Layout of Work Area on Open Bench 5.
Holding Cap and Bulb 5. Waste Beaker 5. Inserting a Pipette in a Pipette Controller 5. Tilting Flasks 5. Boxed Dishes 5. Gassing a Flask 6. Safely Inserting a Pipette into a Pipetting Device 6. Cylinder Clamp 6. Flask for Alcohol Sterilization of Instruments 6.
Microbiological Safety Cabinets 7. Morphology on Feeder Layers 7. Cell Yield and Surface Area 7. Multiwell Plates 7. Petri Dishes 7. Plastic Flasks 7. Multisurface Flask 7. Stirrer Flasks xix Venting Petri Dishes and Flasks 7. Screw-Cap Vials and Flasks 7.
Nonrandom Growth 7. Hollow Fiber Culture 8. Osmometer Effect of Humidity on Temperature in Autoclave Washing and Sterilizing Glassware Sterilizing Capped Bottles Siphon Pipette Washer Washing and Sterilizing Pipettes Semiautomatic Pipette Plugger Bellco Sterilizing Oven Packaging Screw Caps for Sterilization Sterile Filtration Disposable Sterilizing Filters Peristaltic Pump Filtration Large-Scale In-line Filter Assembly Options for Sterile Filtration Reusable Filters Mouse Embryos Mouse Dissection Removing a Chick Embryo from an Egg Options for Primary Culture Primary Explant Culture Warm Trypsin Disaggregation Cell Strainer Cold Trypsin Disaggregation Warm and Cold Trypsinization Dissection of a Chick Embryo Tissue Disaggregation by Collagenase Mechanical Disaggregation Unhealthy Cells Growth Curve and Maintenance He successfully cultivated chicken embryonic cells by using chicken blood plasma, which is readily available, 17 and later successfully cultivated mammalian cells as well.
Carrel is widely believed to be the first person in the world to successfully culture mammalian somatic cells, but the Biographical Memoirs of the National Academy of Sciences contends that the cultivation of guinea pig bone marrow specimens in by Margaret Reed, the future Mrs.
Warren H. Lewis, precedes his success. Lewis and Warren H. Lewis demonstrated that the Locke—Lewis solution—which is modified Locke's solution that additionally contains amino acids, bouillon, and glucose or maltose —is more effective for chick embryo cell cultivation than simple balanced salt solutions. Thus, their research did not lead to the molecular identification of any substance that is essential for cultured cells.
Joilannes P. They typically stop growing after a certain number of divisions ie, the Hayflick limit. In , Wilton R. Gey and his coworkers created an infinitely proliferating human cell line from a tissue of a patient with uterine cervical cancer HeLa cells.
This state of affairs made it easier to examine and to precisely quantify the subtle differences in the effects of culture media on cells.
Thus, the development of culture media advanced rapidly as a result. In order to find the crucial components in those natural materials and to develop defined media that are comparably efficient in the cultivation of cells, relative to the media containing natural ingredients, two main strategies were undertaken.
The first strategy was to use dialyzed serum for the support of cells at minimum levels and to add defined components to maximize the proliferation of cells. The second strategy was not dependent on serum, or even proteins at all, and involved the formulation of media exclusively from definitive components. Neither the publisher nor author shall be liable for any loss of profit or any other commercial damages, including but not limited to special, incidental, consequential, or other damages.
For general information on our other products and services or for technical support, please contact our Customer Care Department within the United States at , outside the United States at or fax Wiley also publishes its books in a variety of electronic formats. Some content that appears in print may not be available in electronic formats. For more information about Wiley products, visit our web site at www. Ian Freshney. Includes index. ISBN cloth 1. F74 Introduction, 1 1.
Historical Background, 1 1. Advantages of Tissue Culture, 6 1. Control of the Environment, 6 1.
Characterization and Homogeneity of Samples, 6 1. Economy, Scale, and Mechanization, 6 1. In vitro Modeling of In vivo Conditions, 7 1.
Limitations, 7 1. Expertise, 7 1. Quantity, 7 1. Dedifferentiation and Selection, 8 1. Origin of Cells, 8 1. Instability, 8 1. Major Differences In vitro, 8 1. Types of Tissue Culture, 8 2. Biology of Cultured Cells, 11 2. The Culture Environment, 11 2. Cell Adhesion, 11 2. Cell Adhesion Molecules, 11 2. Intercellular Junctions, 12 2.
Extracellular Matrix, 13 2. Cytoskeleton, 14 2. Cell Motility, 14 2. Cell Proliferation, 15 2. Cell Cycle, 15 2. Control of Cell Proliferation, 15 2. Differentiation, 16 2. Maintenance of Differentiation, 17 2. Dedifferentiation, 17 2. Cell Signaling, 17 2. Energy Metabolism, 19 2. Origin of Cultured Cells, 20 2. Initiation of the Culture, 21 2. Evolution of Cell Lines, 21 2. Senescence, 22 2. Transformation and the Development of Continuous Cell Lines, 22 3. Laboratory Design, Layout, and Equipment, 25 3.
Layout, Furnishing, and Services, 25 3.