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Culture of animal cells : a manual of basic technique / R. Ian Freshney

Main Author Freshney, R. Ian Country Estados Unidos. Edition 5th ed Publication Hoboken, N.J. : Wiley-Liss, cop. 2005 Description XXVI, 642 p., [24] p. est. : il. ; 29 cm ISBN 0-471-45329-3
978-0-471-45329-1
CDU 57.08 576
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Holdings
Item type Current location Call number Status Date due Barcode Item holds Course reserves
Monografia Biblioteca Prof. Joaquim Pinto Machado
BPM 57.08 - F Available 370239
Monografia Biblioteca Geral da Universidade do Minho
BGUM 57.08 - F Available 371301

Mestrado Integrado em Engenharia Biomédica Laboratórios de Biocompatibilidade e Cultura de Tecidos 2º semestre

Mestrado em Biotecnologia Tecnologia de Cultura de Células 1º semestre

Total holds: 0

Enhanced descriptions from Syndetics:

The most complete resource on the techniques, equipment,principles, and practices of animal cell culture

Since publication of the previous edition of this benchmark text, numerous groundbreaking advances have occurred in stem cell research, cloning, tissue engineering, and in vitro toxicity testing. These and other developments have been incorporated into this fully revised and expanded Fifth Edition of Culture of Animal Cells. In addition, to answer the needs of the exponential increase in newcomers to cell culture, particularly in the biopharmaceutical industry, a completely new chapter on training in cell culture technology has been introduced.

The most complete resource on the techniques, equipment, principles, and practices of animal cell culture, this text offers a complete background related to growth of animal cells in 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 coverage includes:
* An all-new section of training exercises, separated into basic, intermediate, and advanced procedures, cross-referenced to the relevant protocols
* New coverage of stem cells, bioethics, validation, cloning, cell signaling, in vitro toxicity testing, and tissue engineering
* An expanded full-color atlas section, with images of primary culture, cell lines, subculture, differentiation, cancer cells and transformation, three-dimensional culture, contamination, and specialized equipment
* Enhanced treatment of troubleshooting, with full cross-referencing to the relevant protocols and sections of text
* Fully updated references
* The clearest, most consistent presentation of step-by-step protocols available
* Numerous diagrams, photographs, tables, and charts
* Detailed and up-to-date information on reagent preparation and sourcing of materials and equipment, including a fully updated list of suppliers and other resources with Web sites

Indispensable for clinical and biopharmaceutical researchers and scientists, students, trainees, and technicians, this landmark text presents the most accessible and comprehensive introduction available to the culture and experimental manipulation of animal cells.

Table of contents provided by Syndetics

  • Figures (p. xvii)
  • Color Plates (p. xxi)
  • Preface (p. xxiii)
  • Abbreviations (p. xxv)
  • 1 Introduction (p. 1)
  • Background (p. 1)
  • Advantages of Tissue Culture (p. 4)
  • Limitations (p. 5)
  • Major Differences In Vitro (p. 6)
  • Types of Tissue Culture (p. 6)
  • 2 Biology of Cultured Cells (p. 9)
  • The Culture Environment (p. 9)
  • Cell Adhesion (p. 9)
  • Cell Proliferation (p. 11)
  • Differentiation (p. 11)
  • Energy Metabolism (p. 13)
  • Initiation of the Culture (p. 15)
  • Evolution of Cell Lines (p. 15)
  • The Development of Continuous Cell Lines (p. 16)
  • Origin of Cultured Cells (p. 16)
  • 3 Design and Layout (p. 19)
  • Planning (p. 19)
  • Construction and Services (p. 20)
  • Layout (p. 22)
  • 4 Equipment (p. 31)
  • Requirements of a Tissue Culture Laboratory Essential Equipment (p. 31)
  • Beneficial Equipment (p. 40)
  • Useful Additional Equipment (p. 46)
  • Consumable Items (p. 48)
  • 5 Aseptic Technique (p. 51)
  • Objectives of Aseptic Technique (p. 51)
  • Elements of Aseptic Environment (p. 52)
  • Sterile Handling (p. 55)
  • Laminar Flow (p. 56)
  • Standard Procedure (p. 58)
  • Apparatus and Equipment (p. 62)
  • 6 Safety (p. 65)
  • Risk Assessment (p. 65)
  • General Safety (p. 68)
  • Fire (p. 72)
  • Radiation (p. 72)
  • Biohazards (p. 73)
  • 7 Culture Vessels (p. 77)
  • The Substrate (p. 77)
  • Choice of Culture Vessel (p. 78)
  • Specialized Systems (p. 83)
  • Treated Surfaces (p. 83)
  • 8 Media (p. 89)
  • Development of Media (p. 89)
  • Physicochemical Properties (p. 89)
  • Balanced Salt Solutions (p. 93)
  • Complete Media (p. 94)
  • Serum (p. 99)
  • Selection of Medium and Serum (p. 101)
  • Other Supplements (p. 103)
  • 9 Serum-Free Media (p. 104)
  • Disadvantages of Serum (p. 105)
  • Advantages of Serum-Free Media (p. 110)
  • Disadvantages of Serum-Free Media (p. 111)
  • Replacement of Serum (p. 111)
  • Selection of Serum-Free Medium (p. 116)
  • Development of Serum-Free Medium (p. 117)
  • Preparation of Serum-Free Medium (p. 120)
  • Conclusions (p. 120)
  • 10 Preparation and Sterilization (p. 121)
  • Apparatus (p. 121)
  • Reagents and Media (p. 130)
  • Control, Testing, and Storage of Media (p. 146)
  • 11 Primary Culture (p. 149)
  • Types of Primary Cell Culture (p. 149)
  • Isolation of the Tissue (p. 149)
  • Primary Culture (p. 157)
  • 12 Cell Lines (p. 177)
  • Nomenclature (p. 177)
  • Subculture and Propagation (p. 177)
  • Immortalization of Cell Lines (p. 180)
  • Cell Line Designations (p. 180)
  • Selection of Cell Line (p. 181)
  • Routine Maintenance (p. 181)
  • 13 Cloning and Selection (p. 195)
  • Cloning (p. 195)
  • Stimulation of Plating Efficiency (p. 198)
  • Suspension Cloning (p. 200)
  • Isolation of Clones (p. 204)
  • Replica Plating (p. 207)
  • Selective Inhibitors (p. 207)
  • Isolation of Genetic Variants (p. 208)
  • Interaction with Susbstrate (p. 211)
  • 14 Cell Separation (p. 215)
  • Cell Density and Isopyknic Sedimentation (p. 215)
  • Antibody-Based Techniques (p. 218)
  • Magnetic Sorting (p. 219)
  • Cell Size and Sedimentation Velocity (p. 222)
  • Centrifugal Elutriation (p. 222)
  • Fluorescence-Activated Cell Sorting (p. 223)
  • Other Techniques (p. 225)
  • Beginner's Approach to Cell Separation (p. 226)
  • 15 Characterization (p. 229)
  • The Need for Characterization (p. 229)
  • Morphology (p. 231)
  • Chromosome Analysis (p. 241)
  • DNA Content (p. 245)
  • RNA and Protein (p. 249)
  • Enzyme Activity (p. 251)
  • Antigenic Markers (p. 254)
  • Differentiation (p. 257)
  • Authentication (p. 257)
  • 16 Differentiation (p. 259)
  • Expression of In Vivo Phenotype (p. 259)
  • Stages of Commitment and Differentiation (p. 259)
  • Proliferation and Differentiation (p. 260)
  • Commitment and Lineage (p. 260)
  • Markers of Differentiation (p. 261)
  • Induction of Differentiation (p. 262)
  • Differentiation and Malignancy (p. 266)
  • Practical Aspects (p. 266)
  • 17 Transformation (p. 269)
  • Role in Cell Line Characterization (p. 269)
  • What is Transformation? (p. 269)
  • Genetic Instability (p. 269)
  • Immortalization (p. 271)
  • Aberrant Growth Control (p. 277)
  • Tumorigenicity (p. 280)
  • 18 Contamination (p. 285)
  • Sources of Contamination (p. 285)
  • Types of Microbial Contamination (p. 289)
  • Monitoring for Contamination (p. 289)
  • Eradication of Contamination (p. 294)
  • Cross-Contamination (p. 295)
  • Conclusions (p. 296)
  • 19 Cryopreservation (p. 297)
  • Need for Cryopreservation (p. 297)
  • Preservation (p. 297)
  • Cell Banks (p. 307)
  • Transporting Cells (p. 308)
  • 20 Quantitation (p. 309)
  • Cell Counting (p. 309)
  • Cell Weight (p. 314)
  • DNA Content (p. 314)
  • Protein (p. 314)
  • Rates of Synthesis (p. 315)
  • Preparation of Samples for Enzyme Assay and Immunoassay (p. 317)
  • Cytometry (p. 317)
  • Replicate Sampling (p. 318)
  • Cell Proliferation (p. 319)
  • Plating Efficiency (p. 323)
  • Labeling Index (p. 325)
  • Cell Cycle Time (Generation Time) (p. 327)
  • Cell Migration (p. 328)
  • 21 Cytotoxicity (p. 329)
  • Introduction (p. 329)
  • In Vitro Limitations (p. 330)
  • Nature of the Assay (p. 330)
  • Anticancer Drug Screening (p. 340)
  • Transformation (p. 340)
  • Inflammation (p. 343)
  • 22 Specialized Cells (p. 345)
  • Epithelial Cells (p. 345)
  • Mensenchymal Cells (p. 364)
  • Neuroectodermal Cells (p. 373)
  • Hematopoietic Cells (p. 380)
  • Gonads (p. 384)
  • 23 Tumor Cells (p. 385)
  • Sampling (p. 386)
  • Disaggregation (p. 387)
  • Primary Culture (p. 387)
  • Characterization (p. 387)
  • Development of Cell Lines (p. 388)
  • Selective Culture (p. 388)
  • Specific Tumor Types (p. 392)
  • 24 Organotypic Culture (p. 395)
  • Cell Interaction and Phenotypic Expression (p. 395)
  • Organ Culture (p. 396)
  • Histotypic Culture (p. 399)
  • Filter-Well Inserts (p. 403)
  • Cultures of Neuronal Aggregates (p. 405)
  • Organotypic Culture (p. 406)
  • 25 Scale-Up (p. 407)
  • Scale-up in Suspension (p. 407)
  • Scale-up in Monolayer (p. 412)
  • 26 Specialized Techniques (p. 423)
  • Lymphocyte Preparation (p. 423)
  • Autoradiography (p. 424)
  • Time-Lapse Recording (p. 429)
  • Confocal Microscopy (p. 431)
  • Cell Synchrony (p. 431)
  • Culture of Amniocytes (p. 432)
  • Culture of Cells from Poikilotherms (p. 437)
  • 27 Molecular Techniques (p. 443)
  • Molecular Biology in Cell Culture (p. 443)
  • In Situ Molecular Hybridization (p. 443)
  • Somatic Cell Fusion (p. 449)
  • Production of Monoclonal Antibodies (p. 452)
  • DNA Transfer (p. 455)
  • 28 Problem Solving (p. 463)
  • Slow Cell Growth (p. 463)
  • Choice of Medium (p. 465)
  • Unstable Reagents (p. 466)
  • Purity of Constituents (p. 466)
  • Plastics (p. 466)
  • Glassware (p. 466)
  • Microbial Contamination (p. 467)
  • Chemical Contamination (p. 469)
  • Primary Culture (p. 469)
  • Cloning (p. 470)
  • Differentiation (p. 471)
  • Feeding (p. 471)
  • Subculture (p. 471)
  • Cross-Contamination (p. 472)
  • Cryopreservation (p. 472)
  • Granularity of Cells (p. 473)
  • Cell Counting (p. 473)
  • Viability (p. 474)
  • In Conclusion (p. 475)
  • Reagent Appendix (p. 477)
  • Trade Index (p. 483)
  • Glossary (p. 517)
  • References (p. 523)
  • Index (p. 565)

Author notes provided by Syndetics

R. IAN FRESHNEY, PhD, is an honorary senior research fellow in the Centre for Oncology and Applied Pharmacology, part of the Cancer Research UK Beatson Laboratories at the University of Glasgow. He is the author or editor of numerous books and a world-renowned expert on cell culture technique.

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