Animal cell culture:
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Oxford
Oxford Univ. Press
2000
|
| Ausgabe: | 3. ed. |
| Schriftenreihe: | The practical approach series
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XVIII, 315 S. graph. Darst. |
| ISBN: | 0199637970 |
Internformat
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Datensatz im Suchindex
| _version_ | 1804127909855625216 |
|---|---|
| adam_text | Animal Cell Culture
Third Edition
k PiraeitBeaD Approach
Edited by
John R W Masters
3rd Floor Research Laboratories,
University College London
OXFORD
UNIVERSITY PRESS
Contents
List of protocols page xiii
Abbreviations xvii
1 Introduction to basic principles
R Ian Freshney
1 Background 1
2 Biology of cells in culture 2
Origin and characterization 2
Differentiation 3
3 Choice of materials 3
Cell type 3
Source of tissue 4
Subculture 5
Selection of medium 7
Gas phase 8
Culture system 9
4 Procedures 10
Substrate 10
Medium 11
_ Cell culture 11
References - 16
2 Scaling-up of animal cell cultures
Bryan Griffiths
1 Introduction 19
2 General methods and culture parameters
Cell quantification 20
Equipment and reagents 21
Practical considerations 22
Growth kinetics 23
Medium and nutrients 23
pH 24
Oxygen 26
CONTENTS
Types of culture process 30
Summary of factors limiting scale-up 32
3 Monolayer culture 32
Introduction 32
Cell attachment 34
Scaling-up 35
4 Suspension culture 48
Adaptation to suspension culture 49
Static suspension culture 50
Small scale suspension culture 50
Scaling-up factors 53
Stirred bioreactors 54
Continuous-flow culture 56
Airlift fermenter 57
5 Immobilized cultures 58
Immurement cultures 59
Entrapment cultures 62
Porous carriers 62
References 66
3 Cell line preservation and authentication
R J Hay, M M Cleland, S Durkin, and Y A Reid
1 Introduction 69
2 Cell line banking 70
3 Cell freezing and quantitation of recovery 71
Equipment 72
Preparation and freezing 73
Reconstitution and quantitating recovery 74
4 Cell line authentication 78
Species verification 78
Tests for microbial contamination 81
Testing for intraspecies cross-contamination 94
Miscellaneous characterizations and cell line availability 101
Acknowledgements 102
References 102
4 Development of serum-free media
Soverin Karmiol
1 Introduction 105
2 Role of serum and other undefined tissue extracts in cell culture
systems 105-
3 Response curves 106
Proliferating cultures 106
Non-proliferating cultures (hepatocytes) 111
4 Antimicrobials, phenol red, Hepes, and light 112
Phenol red 113
Gentamicin 114
viii
CONTENTS
Hepes 115
Light 116
5 Purity of components 117
6 Fatty acids 117
Acknowledgements 120
References 120
5 Three-dimensional culture
L A Kunz-Schughart and W Mueller-Kleiser
1 Introduction 123
2 Multicellular tumour spheroids (MCTS) 124
MCTS monocultures 124
MCTS co-cultures 133
3 Experimental tissue modelling 139
Current research on tissue modelling 139
Tissue modelling of skin and mucosa 140
Embryoid bodies 143
References 143
6 Tissue engineering
Robert A Brown and Rebecca A Porter
1 Introduction 149
2 Design stages for tissue engineering 150
Tissue engineered skin 152
Tissue engineered urothelium 152
Tissue engineered peripheral nerve implants 153
3 Cell substrates and support materials 154
4 Cell sources 157
5 Orientation 159
Mechanical cues 161
6 Protocols 163
Cell seeding of implantable materials 163
Slow release systems for local control ofTE constructs or repair sites 165
Acknowledgements 168
References 168
7 Cytotoxicity and viability assays
Anne P Wilson
1 Introduction 175
2 Background 176
3 Specific techniques 177
Culture methods 177
Duration of drug exposure and drug concentrations 179
Recovery period 182
ix
CONTENTS
4 End-points 183
Cytotoxicity, viability, and survival 183
Cytotoxicity and viability 183
Survival (reproductive integrity) 191
5 Assay comparisons 192
6 Technical protocols 192
Drugs and drug solutions 192
Drug incubation 194
Assay by survival and proliferative capacity 194
Cytoxicity assays 201
7 Interpretation of results 210
Relationship between cell number and cytoxicity index 210
Dose-response curves 211
8 Pitfalls and troubleshooting 214
Large standard deviations 214
Variation between assays 214
Stimulation to above control levels 214
References 215
8 Fluorescence in situ hybridization
W Nicol Keith
1 Introduction 221
2 Probes 222
3 Probe detection 228
4 A final word on the tricky bits 231
5 FISH resources 232
Solutions 232
Useful books 232
Useful Web sites 232
References 233
9 Genetic modification
Majid Hafezparast _
1 Introduction 235
2 Transfection 235
Calcium phosphate-DNA co-precipitation 237
Lipid-mediated gene transfer (lipofection) 238
Electroporation 240
Staining of cells for expression of 0-galactosidase 243
Rescue of episomal plasmids 244
3 Microcell-mediated chromosome transfer 245
Formation of micronuclei 246
Enucleation 247
Purification of microcells 248
Fusion of microcells to recipient cells 250
Selection of microcell hybrids 250
x
4 Irradiation fusion gene transfer 252
Use in mapping genes 253
Use in positional cloning 254
Irradiation doses 255
References 256
10 Epithelial stem cell identification, isolation, and culture
David Hudson
1 Introduction 259
Clinical application of cultured human stem cells 259
2 Basic principles for identification and purification of stem cells 260
3 Assessment of proliferative heterogeneity 260
4 Methods for the separation of different cell populations 265
Isolation of cells by differential adhesion 266
Separation of cultured and primary cells by flow and immunomagnetic
sorting 270
5 Long-term maintenance of stem cells in culture 273
Keratinocyte stem cells in culture 273
Maintenance of non-epidermal epithelial cells in long-term culture 273
6 Stem cell characterization by immunocytochemistiy 274
Antibody markers of differentiated cell phenotypes 274
Staining cell suspensions using cytospin preparations 277
References 278
11 Senescence, apoptosis, and necrosis
Ian R Kill and Richard G A Faragher
1 Introduction 281
Cellular senescence 281
Cell death 282
Differentiation and de-differentiation 282
2 Simple measures of the population dynamics of primaiy cultures 282
Measurement of the growth fraction 283
Determination of the necrotic fraction of the population 287
Determination of the senescent fraction of the population 288
Determination of the apoptotic fraction of the population using TUNEL 290
3 Other techniques for analysing population dynamics 291
Detection of apoptosis using DNA laddering 292
Inhibition of apoptosis using peptide inhibitors of caspases 293
Determination of the non-dividing fraction of a population by simplified
haptotactic assays, Ponten Plates 295
Determination of telomerase activity and telomere length 296
Acknowledgements 301
References 301
A1 List of suppliers 303
Index 3ii
|
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| dewey-full | 571.6381 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 571 - Physiology & related subjects |
| dewey-raw | 571.6381 |
| dewey-search | 571.6381 |
| dewey-sort | 3571.6381 |
| dewey-tens | 570 - Biology |
| discipline | Biologie |
| edition | 3. ed. |
| format | Book |
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| spelling | Animal cell culture ed. by John R. W. Masters 3. ed. Oxford Oxford Univ. Press 2000 XVIII, 315 S. graph. Darst. txt rdacontent n rdamedia nc rdacarrier The practical approach series Animals Cytology Cell culture Methode (DE-588)4038971-6 gnd rswk-swf Gewebekultur (DE-588)4157245-2 gnd rswk-swf Tierzelle (DE-588)4185509-7 gnd rswk-swf Zellkultur (DE-588)4067547-6 gnd rswk-swf Tiere (DE-588)4060087-7 gnd rswk-swf Zellkultur (DE-588)4067547-6 s Tierzelle (DE-588)4185509-7 s DE-604 Gewebekultur (DE-588)4157245-2 s Tiere (DE-588)4060087-7 s Methode (DE-588)4038971-6 s 1\p DE-604 Masters, John Sonstige oth HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008999772&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 |
| spellingShingle | Animal cell culture Animals Cytology Cell culture Methode (DE-588)4038971-6 gnd Gewebekultur (DE-588)4157245-2 gnd Tierzelle (DE-588)4185509-7 gnd Zellkultur (DE-588)4067547-6 gnd Tiere (DE-588)4060087-7 gnd |
| subject_GND | (DE-588)4038971-6 (DE-588)4157245-2 (DE-588)4185509-7 (DE-588)4067547-6 (DE-588)4060087-7 |
| title | Animal cell culture |
| title_auth | Animal cell culture |
| title_exact_search | Animal cell culture |
| title_full | Animal cell culture ed. by John R. W. Masters |
| title_fullStr | Animal cell culture ed. by John R. W. Masters |
| title_full_unstemmed | Animal cell culture ed. by John R. W. Masters |
| title_short | Animal cell culture |
| title_sort | animal cell culture |
| topic | Animals Cytology Cell culture Methode (DE-588)4038971-6 gnd Gewebekultur (DE-588)4157245-2 gnd Tierzelle (DE-588)4185509-7 gnd Zellkultur (DE-588)4067547-6 gnd Tiere (DE-588)4060087-7 gnd |
| topic_facet | Animals Cytology Cell culture Methode Gewebekultur Tierzelle Zellkultur Tiere |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=008999772&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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