Culture of animal cells: a manual of basic technique
Saved in:
| Main Author: | |
|---|---|
| Format: | Book |
| Language: | English |
| Published: |
New York [u.a.]
Wiley-Liss
1994
|
| Edition: | 3. ed. |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | XXIV, 486 S. Ill., graph. Darst. |
| ISBN: | 0471589667 |
Staff View
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Record in the Search Index
| _version_ | 1804123223180181504 |
|---|---|
| adam_text | CULTURE
OF ANIMAL
CELLS
A Manual of Basic Technique
Third Edition
R IAN FRESHNEY
Department of Medical Oncology
CRC Beatson Laboratories
University of Glasgow
® WILEY-LISS
A JOHN WILEY amp; SONS, INC , PUBLICATION
New York • Chichester • Brisbane • Toronto • Singapore
Contents
Preface to the Third Edition xvii
Preface to the Second Edition xix
Preface to die First Edition xxi
Abbreviations xxiii
1 Introduction
Background 1
Advantages of Tissue Culture 4
Control of the Environment 4
Characterization and Homogeneity of Sample 4
Economy 4
Disadvantages 4
Expertise 4
Quantity 4
DedifFerentiation and Selection S
Origin of Cells 5
Instability 5
Major Differences In Vitro 5
Definition of Types of Tissue Culture 5
2 Biology of the Cultured Cell
The Culture Environment 9
Cell Adhesion 9
Initiation of the Culture 11
Evolution of Cell Lines 11
The Development of Continuous Cell Lines 12
Dedifferentiation 13
What Is a Cultured Cell? 14
Functional Environment 14
Cell Cycle Control 15
Design and Layout of the Laboratory
Planning 17
Construction and Layout 18
Sterile Handling Area 19
Laminar Flow 20
Incubation 21
Hot Room 22
Service Bench 25
Preparation 25
Wash-Up 27
Storage 27
Equipping the Laboratory
Essential Equipment 31
Incubator 31
Sterilizer 31
Refrigerators and Freezers 32
Inverted Microscope 33
Washing Up _ 34
Sterilizing and Drying 36
Water Purification 36
Centrifuge 36
Cell Freezing 36
Beneficial Equipment 38
Laminar Flow 38
Cell Counter 38
Aspiration Pump 38
COz Incubator 39
Balances, pH Meter, and Osmometer 40
Upright Microscope 40
Dissecting Microscope 40
Temperature Recording 40
Magnetic Stirrer 42
Roller Racks 42
Fluid Handling 42
Useful Additional Equipment 46
Low-Temperature Freezer 46
Glassware Washing Machine 46
Video Camera and Monitor 47
Colony Counter 48
Cell Sizing 49
Controlled-Rate Freezer 49
CONTENTS
Centrifugal Elutriator 49
Flow Cytometer 49
Consumable Items 49
Pipettes 49
Culture Vessels 49
Sterile Containers 50
Syringes and Needles 50
Sterilization Filters 50
Aseptic Technique
Objectives of Aseptic Technique 51
Quiet Area 53
Work Surface 53
Personal Hygiene 53
Pipetting 53
Sterile Handling 55
Swabbing 55
Capping 55
Flaming 55
Handling Bottles and Flasks 56
Pouring 56
Laminar Flow 56
Standard Procedure 57
Laboratory Safety and Biohazards
General Safety 62
Operator 62
Equipment 62
Glassware and Sharp Items 63
Chemical Toxicity 63
Gases 63
Liquid Nitrogen 63
Fire 64
Radiation 65
Biohazards 66
Levels of Containment 66
Human Biopsy Material 66
Disposal 68
The Culture Environment: Substrate,
Gas Phase, Medium and Temperature
The Substrate 71
Glass 71
Disposable Plastic 72
Microcarriers 72
viii CONTENTS
Sterilization of Plastics 72
Alternative Artificial Substrates 72
Treated Surfaces 73
Feeder Layers 74
Three-Dimensional Matrices 74
Nonadhesive Substrates 74
Liquid—Gel or Liquid—Liquid Interfaces 75
Hollow Fibers 75
Filter Well Inserts 76
Culture Vessels 76
The Gas Phase 79
Oxygen 79
Carbon Dioxide 80
Media and Supplements 82
Physical Properties 83
pH 83
Buffering 83
Osmolality 83
Temperature 84
Viscosity 84
Surface Tension and Foaming 84
Constituents of Media 84
Balanced Salt Solutions 84
Defined Media 85
Amino Acids 85
Vitamins 88
Salts 88
Glucose 88
Organic Supplements 90
Hormones and Growth Factors 90
Serum 90
Protein 90
Polypeptides 91
Hormones 91
Nutrients and Metabolites 91
Minerals 91
Inhibitors 91
Serum-Free Media 91
Advantages of Serum-Free Medium 92
Selective Media 93
Disadvantages of Serum-Free Media 93
Replacement of Serum 94
Selection and Development of Serum-Free Medium 97
Preparation of Serum-Free Media 98
Commercially Available Serum-Free Media 98
Serum Substitutes 98
Conclusions 98
Selection of Medium and Serum 100
Batch Reservation 100
Testing Serum 100
Other Supplements 101
Embryo Extract 101
Conditioned Medium 101
Incubation Temperature 101
CONTENTS ix
Preparation and Sterilization
Apparatus 10S
Glassware 105
Pipettes 107
Screw Caps 107
Selection of Detergent 109
Miscellaneous Equipment 111
Alternative Methods of Sterilization 112
Reagents and Media 112
Water 112
Balanced Salt Solutions 114
Media 114
Powdered Media 117
Autoclavable Media 117
Filter Sterilization 117
Sterility Testing 118
Culture Testing 119
Storage 121
Serum 122
Preparation and Sterilization of Other Reagents 125
Disaggregation of the Tissue and
Primary Culture
Isolation of the Tissue 127
Mouse Embryos 128
Hen s Egg 129
Human Biopsy Material 133
Primary Culture 133
Primary Expl ant Technique 133
Enzymatic Disaggregation 135
Disaggregation in Warm Trypsin 136
Trypsin at 4°C 136
Chick Embryo Organ Rudiments 139
Other Enzymatic Procedures 142
Collagenase 142
Mechanical Disaggregation 146
Separation of Viable and Nonviable Cells 147
Maintenance of the Culture: Cell Lines
Nomenclature 149
Routine Maintenance 1 SO
Replacement of Medium 150
Volume, Depth, and Surface Area 153
Changing the Medium or Feeding a Culture 153
Maintenance Medium 153
Subculture 153
Cell Concentration and Density at Subculture 156
Propagation in Suspension 156
Slow Cell Growth 157
Cloning and Selection of Specific Cell
Types
Cloning 161
Dilution Cloning 161
Stimulation of Plating Efficiency 162
Multiwell Dishes 166
Semisolid Media 166
Isolation of Clones 169
Suspension Clones 171
Selective Media 172
Isolation of Genetic Variants 172
Interaction With Substrate 173
Selective Adhesion 173
Selective Detachment 174
Nature of Substrate 174
Physical Methods of Cell Separation
Cell Size and Sedimentation Velocity 179
Unit Gravity Sedimentation 179
Centrifugal Elutriation 184
Cell Density and Isopyknic Sedimentation 185
Fluorescence-Activated Cell Sorting 189
Other Techniques 192
Immune Panning Technique for Separation of Human Hemopoietic
Progenitor Cells 192
Magnetic Sorting 194
Characterization
Introduction 197
Species Identification 197
Lineage or Tissue Markers 197
Unique Markers 198
Transformation 200
Morphology 200
Staining 201
Culture Vessels for Cytology: Monolayer Cultures 201
Suspension Culture 201
Photography 204
Chromosome Content 205
Chromosome Preparations • 205
Chromosome Banding 206
Chromosome Analysis 206
Chromosome Painting 207
DNA Content 208
CONTENTS XI
DNA Hybridization 208
DNA Fingerprinting 210
Multilocus DNA Fingerprinting of Cell Lines 210
RNA and Protein 212
Enzyme Activity 213
Isoenzymes 213
Antigenic Markers 215
Indirect Immunofluorescence 216
Differentiation 217
Induction of Differentiation
Stages of Commitment and Differentiation 220
Proliferation and Differentiation 220
Commitment and Lineage 220
Markers of Differentiation 222
Induction of Differentiation 223
Soluble Inducers 223
Cell Interaction 224
Cell—Matrix Interactions 226
Polarity and Cell Shape 226
Differentiation and Malignancy 226
Practical Aspects 227
Preparation of Collagen Gel 227
The Transformed Phenotype
What Is Transformation? 231
Anchorage Independence 234
-Suspension Cloning 234
Contact Inhibition and Density Limitation of Growth 235
Growth on Confluent Monolayers 236
Genetic Abnormalities 238
Chromosomal Aberrations 238
Cell Products and Serum Dependence 238
Tumor Angiogenesis Factor (TAF) 239
Plasminogen Activator 239
Invasiveness 239
Tumorigenesis 239
Contamination
Types of Microbial Contamination 243
Monitoring Cultures for Mycoplasmas 247
Fluorescent Technique for Detecting Mycoplasmas 247
Alternative Methods 249
Eradication of Mycoplasma Infection 249
Detection of Microbial Contamination 250
Persistent Contamination 251
Cross-Contamination 251
Conclusions 251
Instability, Variation, and Preservation
Environment 253
Selective Overgrowth, Transformation, and Senescence 253
Genetic Instability 254
Preservation 254
Selection of Cell Line 254
Standardization of Medium and Serum 255
Cell Freezing 255
Thawing 260
Cell Banks 263
Transporting Cells 264
Quantitation and Experimental Design
Selection of Cell Line 267
Experimental Design 267
Growth Phase - 268
Cell Counting 268
Hemocytometer 268
Electronic Particle Counting 270
Operation of Coulter Cell Counter (Model DI) 272
Cell Sizing 273
Stained Monolayers 273
Cell Weight 274
DNA Content 274
Hoechst 33258 Method 274
DAPI Method 274
Protein 275
Solubilization of Sample 275
Lowry Method 275
Bradford Method 276
Rates of Synthesis 276
Protein Synthesis 276
DNA Synthesis 277
Preparation of Samples for Enzyme Assay and Immunoassay 278
Replicate Sampling 278
Data Analysis 278
Growth Cycle 279
The Lag Phase 280
The Log Phase 281
The Plateau Phase 281
Plating Efficiency 282
Clonal Growth Assay by Dilution Cloning 282
Automatic Colony Counting 282
Labeling Index 282
Growth Fraction 285
Mitotic Index 285
Division Index 285
Cell Cycle Time (Generation Time) 286
Cytometry 286
CONTENTS XIU
Measurement of Viability and
Cytotoxicity
Introduction 287
Limitations of In Vitro Methods 288
Nature of the Assay 288
Short-Term Assays—Viability 288
Long-Term Tests—Survival 290
Microtitration Assays 292
MTT-Based Cytotoxicity Assay 296
Metabolic Tests 299
Drug Interaction 299
Anticancer Drug Screening 299
Predictive Testing 299
Culture Systems 299
Mutagenicity 300
Mutagenesis, Selection of Mutants, and Estimation of Mutant
Frequency 301
Principle of Detecting Mutants and Determining Mutant Frequency 301
Protocol for the L5178Y TK+/~ Mouse Lymphoma Assay 301
Carcinogenicity 306
Inflammation 306
Culture of Specific Cell Types
Epithelial Cells 310
Epidermis 310
Corneal Epithelial Cells: Culture in Serum-Free Medium of Normal Human
Corneal Cells and Cell Lines With Extended Life-Span 313
Breast 315
~ Cervix 316
Gastrointestinal Tract 318
Liver 320
Pancreas 322
Kidney 323
Bronchial and Tracheal Epithelium 325
Prostate 326
Mesenchymal Cells 327
Connective Tissue 327
Adipose Tissue 328
Muscle 328
Cartilage 331
Bone 332
Endothelium 333
Neuroectodermal Cells 335
Neurons 335
Glia 337
Endocrine Cells 341
Melanocytes 341
Hemopoietic Cells 344
Long-Term Bone Marrow Cultures 345
Gonads 348
xiv CONTENTS
Minimal Deviation Tumors 348
Teratomas 348
21 Culture of Tumor Tissue
Sampling 350
Disaggregation 351
Primary Culture 351
Characterization 351
Development of Cell Lines 352
Continuous Cell Lines 352
General Method 353
Selective Culture 353
Selective Media 353
Selective Substrates 354
Confluent Feeder Layers 354
Suspension Cloning 355
Histotypic Culture 355
Spheroids 355
Xenografts 356
Freezing 356
22 Three-Dimensional Culture Systems
Organ Culture 358
Gas and Nutrient Exchange 358
Structural Integrity 359
Growth and Differentiation 359
Limitations of Organ Culture 359
Types of Organ Culture 359
Histotypic Culture 362
Sponge Techniques 363
Hollow Fibers 363
Reaggregation and Spheroids 363
Filter Wells 365
23 Specialized Techniques
Mass Culture Techniques 369
Suspension Culture 369
Monolayer Culture 371
Lymphocyte Preparation 376
Blast Transformation 379
Autoradiography 379
Culture of Cells From Poikilotherms 383
Cell Synchrony 384
Cell Separation 384
Blockade 385
Time-Lapse Recording 385
Confocal Microscopy 387
Culture of Amniocytes 387
Somatic Cell Fusion 389
Selection of Hybrid Clones 391
Gene Transfer 391
Production of Monoclonal Antibodies 394
In Situ Molecular Hybridization 397
Applications and Recent Advances 401
Virus Preparation and Assay 402
In Conclusion 403
Reagent Appendix 405
Trade Index 411
Sources of Materials 411
Addresses of Commercial Suppliers 417
Scientific Societies With Interests in Tissue Culture
Glossary 433
References 437
General Textbooks for Further Reading 468
Useful Journals 468
Index 471
|
| any_adam_object | 1 |
| author | Freshney, Robert Ian 1938-2019 |
| author_GND | (DE-588)1055981578 |
| author_facet | Freshney, Robert Ian 1938-2019 |
| author_role | aut |
| author_sort | Freshney, Robert Ian 1938-2019 |
| author_variant | r i f ri rif |
| building | Verbundindex |
| bvnumber | BV008870181 |
| callnumber-first | Q - Science |
| callnumber-label | QH585 |
| callnumber-raw | QH585.2 |
| callnumber-search | QH585.2 |
| callnumber-sort | QH 3585.2 |
| callnumber-subject | QH - Natural History and Biology |
| classification_rvk | WX 6600 WX 6603 |
| classification_tum | BIO 770f BIO 655f CIT 972f BIO 768f |
| ctrlnum | (OCoLC)299764373 (DE-599)BVBBV008870181 |
| dewey-full | 591.0724 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 591 - Specific topics in natural history of animals |
| dewey-raw | 591.0724 |
| dewey-search | 591.0724 |
| dewey-sort | 3591.0724 |
| dewey-tens | 590 - Animals |
| discipline | Biologie Chemie-Ingenieurwesen Biotechnologie |
| edition | 3. ed. |
| format | Book |
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| id | DE-604.BV008870181 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T17:26:21Z |
| institution | BVB |
| isbn | 0471589667 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-005865844 |
| oclc_num | 299764373 |
| open_access_boolean | |
| owner | DE-20 DE-29T DE-29 DE-M49 DE-BY-TUM DE-11 DE-188 DE-578 |
| owner_facet | DE-20 DE-29T DE-29 DE-M49 DE-BY-TUM DE-11 DE-188 DE-578 |
| physical | XXIV, 486 S. Ill., graph. Darst. |
| publishDate | 1994 |
| publishDateSearch | 1994 |
| publishDateSort | 1994 |
| publisher | Wiley-Liss |
| record_format | marc |
| spelling | Freshney, Robert Ian 1938-2019 Verfasser (DE-588)1055981578 aut Culture of animal cells a manual of basic technique R. Ian Freshney 3. ed. New York [u.a.] Wiley-Liss 1994 XXIV, 486 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Cell Culture cabt Gewebekultur (DE-588)4157245-2 gnd rswk-swf Tiere (DE-588)4060087-7 gnd rswk-swf Zellkultur (DE-588)4067547-6 gnd rswk-swf Tierzelle (DE-588)4185509-7 gnd rswk-swf Methode (DE-588)4038971-6 gnd rswk-swf Gewebekultur (DE-588)4157245-2 s Tierzelle (DE-588)4185509-7 s DE-604 Zellkultur (DE-588)4067547-6 s Tiere (DE-588)4060087-7 s Methode (DE-588)4038971-6 s 1\p DE-604 2\p DE-604 HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=005865844&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk 2\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk |
| spellingShingle | Freshney, Robert Ian 1938-2019 Culture of animal cells a manual of basic technique Cell Culture cabt Gewebekultur (DE-588)4157245-2 gnd Tiere (DE-588)4060087-7 gnd Zellkultur (DE-588)4067547-6 gnd Tierzelle (DE-588)4185509-7 gnd Methode (DE-588)4038971-6 gnd |
| subject_GND | (DE-588)4157245-2 (DE-588)4060087-7 (DE-588)4067547-6 (DE-588)4185509-7 (DE-588)4038971-6 |
| title | Culture of animal cells a manual of basic technique |
| title_auth | Culture of animal cells a manual of basic technique |
| title_exact_search | Culture of animal cells a manual of basic technique |
| title_full | Culture of animal cells a manual of basic technique R. Ian Freshney |
| title_fullStr | Culture of animal cells a manual of basic technique R. Ian Freshney |
| title_full_unstemmed | Culture of animal cells a manual of basic technique R. Ian Freshney |
| title_short | Culture of animal cells |
| title_sort | culture of animal cells a manual of basic technique |
| title_sub | a manual of basic technique |
| topic | Cell Culture cabt Gewebekultur (DE-588)4157245-2 gnd Tiere (DE-588)4060087-7 gnd Zellkultur (DE-588)4067547-6 gnd Tierzelle (DE-588)4185509-7 gnd Methode (DE-588)4038971-6 gnd |
| topic_facet | Cell Culture Gewebekultur Tiere Zellkultur Tierzelle Methode |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=005865844&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT freshneyrobertian cultureofanimalcellsamanualofbasictechnique |