Cell and tissue culture for medical research:
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Chichester [u.a.]
Wiley
2000
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XXII, 443 S. graph. Darst. |
| ISBN: | 0471852139 |
Internformat
MARC
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| 264 | 1 | |a Chichester [u.a.] |b Wiley |c 2000 | |
| 300 | |a XXII, 443 S. |b graph. Darst. | ||
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Datensatz im Suchindex
| _version_ | 1804128124354428928 |
|---|---|
| adam_text | Contents
Contributors xv
Preface xxiii
Safety xxiv
CHAPTER 1 BASIC LABORATORY SET UP AND PROCEDURES 1
1.1 General Introduction 3
References 4
1.2 Cryopreservation: Basic Techniques and Storage 5
Basic techniques 5
procedure: Cryopreservation using controlled rate
freezing apparatus 6
Long term storage 7
supplementary procedure: Thawing 8
alternative procedure: Mechanical freezing and
two stage freezing 8
Discussion 9
Literature review 10
References 11
1.3 Haemocytometer Cell Counts and Viability Studies 12
procedure: Haemocytometer cell count 12
Discussion 16
1.4 Prophylactic Use of Antibiotics in Cells and Tissues
with a High Risk of Microbial Contamination 17
procedure: Determining the cytotoxicity of
antibiotics in cell and tissue culture 19
alternative procedure: Cytotoxicity test in
lymphoblastoid cell cultures 20
Stability of antibiotics in cell and tissue culture medium 21
Antibiotic assays 21
preliminary procedure: Antibiotic solution 22
preliminary procedure: Preparation of assay plates 22
procedure: Diffusion assay 22
Discussion 24
References 25
1.5 Enzymic Techniques for Tissue Dissociation 27
Tissue types 28
Enzymes 29
Preparation and storage of enzymes 31
Discussion: Optimization of cell dissociation 32
References 36
vi CELL AND TISSUE CULTURE FOR MEDICAL RESEARCH
1.6 Routine Subculturing 38
General 38
procedure: Subculture attached cells 39
alternative procedure: Serum free cultures 40
alternative procedure: Mechanical removal of cells 40
Discussion: Alternative Proteases 41
procedure: Subculture suspension cells 42
alternative procedure: Subculture by dilution 43
Discussion: Serum free cultures 43
Reference 44
CHAPTER 2 SPECIALIZED TECHNIQUES 45
2.1 Cell Quantification An Overview 47
Haemocytometer cell counts 48
Colony forming efficiency 48
MTT assay 49
LDH activity 49
Neutral Red (NR) assay 49
Choice of method 49
References 50
Background reading 50
2.2 Mycoplasma 51
A Detection of Mycoplasma 51
procedure: DNA stain 52
alternative procedure: Use of indicator cell lines 54
supplementary procedure: Microbiological culture 55
supplementary procedure: Elimination of
contamination 56
Discussion 56
References 57
B Detection of Mycoplasma by DNA Amplification (PCR) 58
procedure: Amplification 59
Discussion 60
References 61
2.3 Bacteria and Fungi 62
procedure: Detection of bacteria and fungi in cell
cultures 62
Discussion 64
References 64
2.4 Elimination of Contamination 65
procedure: Eradication 65
Discussion 65
References 67
2.5 Removal of Fibroblasts 68
Choice of method 68
preliminary procedure: Preparation of dishes for
panning 69
CONTENTS vii
preliminary procedure: Preparation of cells for
panning 69
procedure: Panning 70
alternative procedure: Complement and
antibodies 70
rapid procedure: EDTA/trituration 70
supplementary procedure: Other techniques 71
supplementary procedure: Immunoological techniques 72
Literature review 73
References 73
2.6 Identity Testing An Overview 75
Cytogenic analysis 75
Isoenzyme analysis 76
DNA fingerprinting and DNA profiling 76
References 77
2.7 Scale up of Laboratory Cultures 79
procedure: Roller Bottle Culture 79
supplementary procedure: Extended Surface
Area Roller Bottle (ESRB) 80
supplementary procedure: Spinner flask culture 81
supplementary procedure: Microcarrier culture 83
preliminary procedure: Siliconization 86
procedure: Microcarrier culture 87
Discussion 88
References 89
Background reading 90
CHAPTER 3 CELL CHARACTERIZATION AND ANALYSIS 91
3.1 Techniques for Cell Characterization and Analysis
An Overview 93
3.2 Image Cytometry An Overview 95
Goals of medical microscopic imaging 95
Diversity of microscopic imaging 95
Laboratory requirements 98
Major ergonomic weakness 99
Comment 99
33 Fluorescent Image Cytometry 101
procedure: Image acquisition 102
Discussion: Critical parameters 104
References 104
34 Immunocytochemical Labelling and Analysis for
• Light Microscopy 106
Cell culture 106
Detection method 107
¦ Alternative detection methods 114
Controls 114
References 114
viii CELL AND TISSUE CULTURE FOR MEDICAL RESEARCH
3.5 Flow Cytometry An Overview 115
Instrumentation 115
Data analysis 117
Cell sorting 119
Applications 119
Choice of instrument 121
Further reading 121
3.6 Electron Microscopy An Overview 122
Instrumentation 122
Preparation techniques 128
References 137
3.7 Cell Death in Culture Systems (Kinetics of Cell Death) 139
procedure: Morphological characterization of cell
death 140
procedure: Biochemical characterization of cell death 140
supplementary procedure: Purification of
apoptotic cells 144
Discussion 144
References 145
CHAPTER 4 CELL LINE DERIVATION METHODS 147
4.1 Cell Line Derivation An Overview 149
Human cell lines 152
Risk assessment 152
Reference 153
4.2 Cell Lines for Medical Research 154
Introduction 154
VERO 155
A2780 156
NCI H460 156
NCI H596 156
MRC 5 157
MCF 7 157
FDCP mix clone A4 158
HIT T15 158
C6 Glial Cells 158
SK Mel 1 159
WiDr 159
K 562 159
A6 160
HUV EC C 160
PtK2 (NBL 5) 161
References 162
4.3 Explant Cultures 165
procedure: Preparation of human tissue explants 165
procedure: Preparation of animal tissue explants 167
Culture vessels 169
CONTENTS ix
Procedure A 169
Procedure B 170
Discussion 170
References 171
4.4 Haemopoetic Cells in Culture: Human B and T
Lymphocytes 172
Introduction 172
A EBV transformation 173
procedure: Separation of mononuclear cells from
peripheral blood 173
supplementary procedure: Preparation of
EBV containing supernatant 174
supplementary procedure: Titration of EBV
supernatant using cord blood mononuclear cells 175
procedure: Establishment of human B lymphoblastoid
cell lines from peripheral blood mononuclear cells 176
References 178
B Cryopreservation of human blood for B cell
immortalization 179
procedure: Cryopreservation and immortalization
procedure 181
Discussion 184
References 186
C T cells and thymocytes 187
procedure: Sources of T cells 188
procedure: Procedures for establishing T cells
reactive with exogenous soluble antigen 189
procedure: Assay for the antigen specificity of
T cells reactive with exogenous soluble antigen 192
supplementary procedure: Cloning of T cells
reactive with exogenous soluble antigen 193
supplementary procedure: Improved method for
the expansion of human T cell clones 194
procedure: Procedures for establishing T cells
reactive with endogenous cell surface antigen 196
procedure: Assay for the antigen specificity of
T cells reactive with cell tissue surface antigen 197
supplementary procedure: Cloning of T cells
reactive with endogenous cell surface antigen 198
procedure: Mass long term cultures of human T cells 199
supplementary procedure: Mass cultures of
CD4+ and CD8+ cells 201
Discussion 202
References 203
4 5 Establishment of Human Leukaemia Cell Lines 205
Establishment of human leukaemia cell lines: Basic
protocol 205
x CELL AND TISSUE CULTURE FOR MEDICAL RESEARCH
Maintenance of human leukaemia cell lines: Basic
protocol 210
Storage of human leukaemia cell lines: Basic protocol 213
Commentary 215
Background reading 218
4.6 Derivation and Subculturing of Human Diploid Cell
Strains 219
procedure: Derivation and subculturing 219
Theory of population increase by subcultivation 222
References 223
4.7 Rapid Initiation of Adherent Monolayer Cell Cultures:
The Cold Cocktail Method 224
Introduction 224
Materials 225
Reagents, solutions and media 226
Procedure 227
Troubleshooting 231
Alternative protocols 231
Background 233
Acknowledgement 234
References 234
4.8 Cultures of Human Keratinocytes 235
procedure: Skin biopsies 235
procedure: Explant culture 236
procedure: Dispersed epidermal cell culture 237
procedure: Reconstructed epidermis 240
References 242
Background reading 244
4.9 The Isolation and Maintenance of Human Gastric
Epithelial Cells in Primary Culture 245
preliminary procedure: Gastric epithelial cell
adhesion requirements 245
procedure: Processing a human gastric biopsy sample 248
supplementary procedure: Dissociation of human
gastric epithelial cells 248
supplementary procedure: Maintenance of human
gastric epithelium in long term culture 249
supplementary procedure: Propagation of cells 249
Discussion 250
Acknowledgements 250
References 250
4.10 Replicative and Functional Cultures of Normal Human
Hepatocytes 251
Introduction 251
Equipment and material 252
Basic procedure 253
Characterization of hepatic function 260
CONTENTS xi
Discussion 264
References 266
4.11 Embryonic Kidney in Organ Culture 269
preliminary procedure: The preparation of
Nuclepore filters and pancreatin trypsin 270
preliminary procedure: Pre treatment of
Nuclepore filters 271
procedure: Setting up the cultures 271
Discussion 274
References 274
4.12 The Culture of Amniocytes, Chorionic Villi, and
Human Foetal and Placental Tissue 275
procedure: Culture of amniocytes 277
procedure: Culture of human foetal and placental
tissues for chromosome analysis 280
procedure: Culture of chorionic villi for prenatal
chromosome analysis 283
alternative procedure: In situ amniocyte cell
culture and harvesting 286
Discussion 288
References 290
4.13 Endothelial Cells (from the Umbilical Cord Vein) 291
procedure: Endothelial cell extraction 293
Cell cultures 294
Complementary Techniques 296
supplementary procedure: Culture dish
coating 296
supplementary procedure: von Willebrand factor
characterization 297
Conclusion 298
References 298
4.14 Dissociated Nerve Cells 299
procedure: Culture of dissociated nerve cells plated
on poly lysine coated glass coverslips or smooth plastic
surfaces 299
alternative procedure: Culture of dissociated
nerve cells on uncoated microporous membranes 303
Discussion 303
References 304
4.15 Neuronal and Glial Tumours IN VITRO An Overview 306
Introduction 306
Neuronal neoplasms 307
Glial neoplasms 307
Embryonal neoplasms 308
Growth of nervous system tumours in vitro 309
Nervous system tumours in culture 310
References 317
xii CELL AND TISSUE CULTURE FOR MEDICAL RESEARCH
4.16 Colon Adenocarcinoma Cells (Primary/established
Cultures of Human Colon Carcinomas) 321
preliminary procedure: Preparation of
collagen coated flasks 321
preliminary procedure: Preparation of Swiss
3T3 feeder cells 322
procedure: Establishment of cell lines from
colorectal tumours 322
Discussion 326
References 328
CHAPTER 5 MODERN CLINICAL APPLICATIONS OF
CULTURED CELLS 329
5.1 Immortalization Methods An Overview 331
References 332
5.2 Long term Maintenance and Stability of Hybridomas 334
procedure: Endpoint cloning 334
alternative procedure: Mini cloning 336
alternative procedure: Soft agar cloning 336
procedure: Long term maintenance 338
alternative procedure: Monitoring of antibody
secretion using quantitative ELISA 341
5.3 Organic Culture Techniques in Tissue Banking 345
Corneal structure and function 345
Corneal transplantation 346
Corneal storage by organ culture 346
Graft outcome 349
Future developments 349
References 350
5.4 Cell and Tissue Engineering 352
Three dimensional cell culture systems 352
Tissue engineered skin 357
Applications of tissue engineering 357
Current advances 358
References 360
5.5 Gene Therapy and Recombinant Cells 364
A Recombinant DNA cell lines 364
Overview 364
References 367
B Adenovirus vectors 370
The adenovirus genome 370
Adenoviruses as expression vectors 373
Vector construction and rescue 374
procedure: Infection of cells with adenovirus and
isolation of adenovirus DNA from infected cells 376
procedure: Rescue of adenovirus early replacement
vectors 378
CONTENTS xiii
Discussion 385
References 385
C Retroviruses and retroviral vectors 388
References 392
D Retroviral vector mediated gene transfer 393
Selection of retrovirus helper cells and design of
retroviral vectors 393
Construction of retroviral vectors with inserted
gene sequences 395
procedure: Transfection of mammalian cells with
retroviral vectors 396
procedure: Generation of helper cells stably
producing a relatively homogeneous supply of vector
virus 398
supplementary procedure: Infection of susceptible
target cells with vector virus produced from helper
cell lines 399
References 400
5.6 In Vitro Toxicity Testing 402
Toxicity of xenobiotics 402
The use of cultured cells in pharmaco toxicological
research 402
The choice of the in vitro biological model: cell lines
versus primary cultures 403
Toxicity risk assessment in cultured cells 404
Applications of in vitro methods in biomedical
toxicology 418
References 419
APPENDIX 1 TERMINOLOGY 425
Some aspects of the problem 425
Solutions to the problem 426
Terminology associated with cell, tissue and organ
culture, molecular biology and molecular genetics 426
References 433
APPENDIX 2 COMPANY ADDRESSES 434
INDEX 439
|
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| illustrated | Illustrated |
| indexdate | 2024-07-09T18:44:15Z |
| institution | BVB |
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| language | English |
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| spellingShingle | Cell and tissue culture for medical research Tissue Culture cabt Cell Culture cabt Celkweek gtt Cell culture Cellules - Culture - Manuels de laboratoire Medicine - Research Onderzoek gtt Tissue culture Tissus (Histologie) - Culture - Manuels de laboratoire Weefselculturen gtt Medizin Cell Culture Techniques Laboratory Manuals Cell culture Laboratory manuals Culture Techniques Laboratory Manuals Tissue culture Laboratory manuals Zellkultur (DE-588)4067547-6 gnd Labormedizin (DE-588)4073931-4 gnd Gewebekultur (DE-588)4157245-2 gnd |
| subject_GND | (DE-588)4067547-6 (DE-588)4073931-4 (DE-588)4157245-2 |
| title | Cell and tissue culture for medical research |
| title_auth | Cell and tissue culture for medical research |
| title_exact_search | Cell and tissue culture for medical research |
| title_full | Cell and tissue culture for medical research ed. by Alan Doyle ... |
| title_fullStr | Cell and tissue culture for medical research ed. by Alan Doyle ... |
| title_full_unstemmed | Cell and tissue culture for medical research ed. by Alan Doyle ... |
| title_short | Cell and tissue culture for medical research |
| title_sort | cell and tissue culture for medical research |
| topic | Tissue Culture cabt Cell Culture cabt Celkweek gtt Cell culture Cellules - Culture - Manuels de laboratoire Medicine - Research Onderzoek gtt Tissue culture Tissus (Histologie) - Culture - Manuels de laboratoire Weefselculturen gtt Medizin Cell Culture Techniques Laboratory Manuals Cell culture Laboratory manuals Culture Techniques Laboratory Manuals Tissue culture Laboratory manuals Zellkultur (DE-588)4067547-6 gnd Labormedizin (DE-588)4073931-4 gnd Gewebekultur (DE-588)4157245-2 gnd |
| topic_facet | Tissue Culture Cell Culture Celkweek Cell culture Cellules - Culture - Manuels de laboratoire Medicine - Research Onderzoek Tissue culture Tissus (Histologie) - Culture - Manuels de laboratoire Weefselculturen Medizin Cell Culture Techniques Laboratory Manuals Cell culture Laboratory manuals Culture Techniques Laboratory Manuals Tissue culture Laboratory manuals Zellkultur Labormedizin Gewebekultur |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=009103721&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT doylealan cellandtissuecultureformedicalresearch |