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Culture of animal cells : a manual of basic technique / R. Ian Freshney.
Holman Biotech Commons Oversize QH585.2 .F74 2000
Available
LIBRA QH585.2 .F74 2000
Available from offsite location
- Format:
- Book
- Author/Creator:
- Freshney, R. Ian.
- Language:
- English
- Subjects (All):
- Tissue culture--Laboratory manuals.
- Tissue culture.
- Cell culture--Laboratory manuals.
- Cell culture.
- Cells, Cultured.
- Tissue Culture.
- Medical Subjects:
- Cells, Cultured.
- Tissue Culture.
- Genre:
- Laboratory manuals.
- Physical Description:
- xxvi, 577 pages, 12 unnumbered pages of plates : illustrations (some color) ; 29 cm
- Edition:
- Fourth edition.
- Place of Publication:
- New York : Wiley-Liss, 2000.
- Summary:
- A vital training manual of the fundamental principles and techniques of animal cell culture. Crucial updates have been made in this edition to include advances in the fields of biology, media, specialized techniques, biotechnology, DNA transfer, tumor culture, references, and suppliers.
- Contents:
- Advantages of Tissue Culture 4
- Limitations 5
- Major Differences In Vitro 6
- Types of Tissue Culture 6
- 2 Biology of Cultured Cells 9
- The Culture Environment 9
- Cell Adhesion 9
- Cell Proliferation 11
- Differentiation 11
- Energy Metabolism 13
- Initiation of the Culture 15
- Evolution of Cell Lines 15
- The Development of Continuous Cell Lines 16
- Origin of Cultured Cells 16
- 3 Design and Layout 19
- Planning 19
- Construction and Services 20
- Layout 22
- 4 Equipment 31
- Requirements of a Tissue Culture Laboratory Essential Equipment 31
- Beneficial Equipment 40
- Consumable Items 48
- 5 Aseptic Technique 51
- Objectives of Aseptic Technique 51
- Elements of Aseptic Environment 52
- Sterile Handling 55
- Laminar Flow 56
- Standard Procedure 58
- Apparatus and Equipment 62
- 6 Safety 65
- Risk Assessment 65
- General Safety 68
- Fire 72
- Radiation 72
- Biohazards 73
- 7 Culture Vessels 77
- The Substrate 77
- Choice of Culture Vessel 78
- Specialized Systems 83
- Treated Surfaces 83
- 8 Media 89
- Development of Media 89
- Physicochemical Properties 89
- Balanced Salt Solutions 93
- Complete Media 94
- Serum 99
- Selection of Medium and Serum 101
- Other Supplements 103
- 9 Serum-Free Media 104
- Disadvantages of Serum 105
- Advantages of Serum-Free Media 110
- Disadvantages of Serum-Free Media 111
- Replacement of Serum 111
- Selection of Serum-Free Medium 116
- Development of Serum-Free Medium 117
- Preparation of Serum-Free Medium 120
- 10 Preparation and Sterilization 121
- Apparatus 121
- Reagents and Media 130
- Control, Testing, and Storage of Media 146
- 11 Primary Culture 149
- Types of Primary Cell Culture 149
- Isolation of the Tissue 149
- Primary Culture 157
- 12 Cell Lines 177
- Nomenclature 177
- Subculture and Propagation 177
- Immortalization of Cell Lines 180
- Cell Line Designations 180
- Selection of Cell Line 181
- Routine Maintenance 181
- 13 Cloning and Selection 195
- Cloning 195
- Stimulation of Plating Efficiency 198
- Suspension Cloning 200
- Isolation of Clones 204
- Replica Plating 207
- Selective Inhibitors 207
- Isolation of Genetic Variants 208
- Interaction with Susbstrate 211
- 14 Cell Separation 215
- Cell Density and Isopyknic Sedimentation 215
- Antibody-Based Techniques 218
- Magnetic Sorting 219
- Cell Size and Sedimentation Velocity 222
- Centrifugal Elutriation 222
- Fluorescence-Activated Cell Sorting 223
- Other Techniques 225
- Beginner's Approach to Cell Separation 226
- 15 Characterization 229
- The Need for Characterization 229
- Morphology 231
- Chromosome Analysis 241
- DNA Content 245
- RNA and Protein 249
- Enzyme Activity 251
- Antigenic Markers 254
- Differentiation 257
- Authentication 257
- 16 Differentiation 259
- Expression of In Vivo Phenotype 259
- Stages of Commitment and Differentiation 259
- Proliferation and Differentiation 260
- Commitment and Lineage 260
- Markers of Differentiation 261
- Induction of Differentiation 262
- Differentiation and Malignancy 266
- Practical Aspects 266
- 17 Transformation 269
- Role in Cell Line Characterization 269
- What is Transformation? 269
- Genetic Instability 269
- Immortalization 271
- Aberrant Growth Control 277
- Tumorigenicity 280
- 18 Contamination 285
- Sources of Contamination 285
- Types of Microbial Contamination 289
- Monitoring for Contamination 289
- Eradication of Contamination 294
- Cross-Contamination 295
- 19 Cryopreservation 297
- Need for Cryopreservation 297
- Preservation 297
- Cell Banks 307
- Transporting Cells 308
- 20 Quantitation 309
- Cell Counting 309
- Cell Weight 314
- DNA Content 314
- Protein 314
- Rates of Synthesis 315
- Preparation of Samples for Enzyme Assay and Immunoassay 317
- Cytometry 317
- Replicate Sampling 318
- Cell Proliferation 319
- Plating Efficiency 323
- Labeling Index 325
- Cell Cycle Time (Generation Time) 327
- Cell Migration 328
- 21 Cytotoxicity 329
- In Vitro Limitations 330
- Nature of the Assay 330
- Anticancer Drug Screening 340
- Transformation 340
- Inflammation 343
- 22 Specialized Cells 345
- Epithelial Cells 345
- Mensenchymal Cells 364
- Neuroectodermal Cells 373
- Hematopoietic Cells 380
- Gonads 384
- 23 Tumor Cells 385
- Sampling 386
- Disaggregation 387
- Primary Culture 387
- Characterization 387
- Development of Cell Lines 388
- Selective Culture 388
- Specific Tumor Types 392
- 24 Organotypic Culture 395
- Cell Interaction and Phenotypic Expression 395
- Organ Culture 396
- Histotypic Culture 399
- Filter-Well Inserts 403
- Cultures of Neuronal Aggregates 405
- Organotypic Culture 406
- 25 Scale-Up 407
- Scale-up in Suspension 407
- Scale-up in Monolayer 412
- 26 Specialized Techniques 423
- Lymphocyte Preparation 423
- Autoradiography 424
- Time-Lapse Recording 429
- Confocal Microscopy 431
- Cell Synchrony 431
- Culture of Amniocytes 432
- Culture of Cells from Poikilotherms 437
- 27 Molecular Techniques 443
- Molecular Biology in Cell Culture 443
- In Situ Molecular Hybridization 443
- Somatic Cell Fusion 449
- Production of Monoclonal Antibodies 452
- DNA Transfer 455
- 28 Problem Solving 463
- Slow Cell Growth 463
- Choice of Medium 465
- Unstable Reagents 466
- Purity of Constituents 466
- Plastics 466
- Glassware 466
- Microbial Contamination 467
- Chemical Contamination 469
- Primary Culture 469
- Cloning 470
- Differentiation 471
- Feeding 471
- Subculture 471
- Cross-Contamination 472
- Cryopreservation 472
- Granularity of Cells 473
- Cell Counting 473
- Viability 474.
- Notes:
- Includes bibliographical references and index.
- Local Notes:
- Acquired for the Penn Libraries with assistance from the Class of 1932 Fund.
- ISBN:
- 0471348899
- OCLC:
- 41266026
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