Cell growth and apoptosis: a practical approach
This Practical Approach volume is unique in describing the key concepts of cell growth and cell death, particularly by apoptosis, and in providing detailed methods to study these processes. The chapters are written by experts in the field and cover a wide range of important subject areas including c...
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Oxford
IRL Press
1995
|
| Schriftenreihe: | The practical approach series
159 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Zusammenfassung: | This Practical Approach volume is unique in describing the key concepts of cell growth and cell death, particularly by apoptosis, and in providing detailed methods to study these processes. The chapters are written by experts in the field and cover a wide range of important subject areas including cell-cycle traverse, chromosomal abnormalities detected at cell division, DNA damage, and cell death or senescence. The establishment of cell lines and optimization of cell proliferation by growth factors are also described. The result is an invaluable laboratory manual for studies of growth and its abnormalities in mammalian cells. Cell Growth and Apoptosis: A Practical Approach provides state-of-the-art protocols for studies of normal and abnormal cell growth, including cancer and other cell-proliferation disorders, together with a critical appraisal of the various methodologies in use. Its coverage of human cells will make it particularly useful for health professionals. |
| Beschreibung: | XXVI, 269 S. Ill., graph. Darst. |
| ISBN: | 0199635692 0199635684 |
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| 490 | 1 | |a The practical approach series |v 159 | |
| 520 | 3 | |a This Practical Approach volume is unique in describing the key concepts of cell growth and cell death, particularly by apoptosis, and in providing detailed methods to study these processes. The chapters are written by experts in the field and cover a wide range of important subject areas including cell-cycle traverse, chromosomal abnormalities detected at cell division, DNA damage, and cell death or senescence. The establishment of cell lines and optimization of cell proliferation by growth factors are also described. The result is an invaluable laboratory manual for studies of growth and its abnormalities in mammalian cells. Cell Growth and Apoptosis: A Practical Approach provides state-of-the-art protocols for studies of normal and abnormal cell growth, including cancer and other cell-proliferation disorders, together with a critical appraisal of the various methodologies in use. Its coverage of human cells will make it particularly useful for health professionals. | |
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Datensatz im Suchindex
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|---|---|
| adam_text | Cell Growth and
Apoptosis
A Practical Approach
Edited by
GEORGE P STUDZINSKI
Department of Laboratory Medicine and Pathology,
UMDNJ-New Jersey Medical School,
Newark, NJ
OXFORD UNIVERSITY PRESS
Oxford New York Tokyo
Contents
Last of contributors xix
Abbreviations xxm
1 Measuring parameters of growth 1
Renato Baserga
1 Introduction 1
2 Growth parameters 2
Indirect measurements of cell number 3
DNA amount 3
RNA amount 4
Protein amount 4
3 Mitoses 4
4 DNA synthesis 7
Autoradiography 7
5 Determination of parameters of growth by
autoradiography with [3H]thymidine 12
Growth fraction 12
Quiescent cells and stationary cell populations 13
Phases of the cell cycle 14
Short method for growth fraction and cell cycle phases IS
Flow of cells through the cell cycle 16
6 Quiescence of cells 17
References 18
2 Cell cycle analysis by flow cytometry 2i
Harry A Crissman
1 Introduction 21
2 Cell preparation and fixation 22
3 DNA-specific fluorochromes commonly used for staining 23
DNA reactive Hoechst dyes, DAPI and DIPI 24
Mithramycin, chromomycin, and olivomycin 24
Propidium iodide (PI), ethidium bromide (EB), 7-amino-
actinomycin D, and hydroethidine (HE) 24
Acridine Orange 25
TOTO-1, YOYO-1, TO-PRO-1, and YO-PRO-1 25
Contents
} 4 Cell staining 25
Viable cell staining with Hoechst 33342 25
Staining of fixed cells and unfixed nuclei 26
5 Instrumentation setup and analysis 27
Standards 27
Application 28
6 Simultaneous DNA and protein assay 28
Cell staining 29
Instrument setup and analysis 29
Applications 30
7 Cellular bromodeoxyuridine and DNA contents 30
Differential fluorescence analysis of BrdUrd content 32
BrdUrd pulse labelling 32
Cell staining with Hoechst 33342 (HO) and mithramycin (MI) 33
Instrument setup for differential fluorescence analysis 34
Analysis of stained cells 34
8 Correlated, three-colour, DNA-RNA-protein determinations 36
Cell staining 37
Three-laser FCM setup for analysis of DNA-RNA-protein 37
Analysis of three-colour stained cells 38
Controls for optimal colour resolution 39
9 Correlated analysis of cyclin Bl and cell cycle 40
Cell staining 40
Analysis by flow cytometry 41
Acknowledgements 42
References • 42
3 Analysis of flow cytometric DNA histograms 45
Peter S Rabinovitch
1 Introduction 45
2 Histogram analysis 45
3 Cell cycle modelling 48
The DNA content cell cycle 48
Model components 50
Debris modelling 51
Aggregation modelling 53
Complexity of the model 54
4 Reliability of histogram analysis results 56
CV and peak shape 56
Quantitation of aggregates and debris 56
Proportion of aneuploid cells 57
Contents
Statistical measures derived from the fitting model 57
Sensitivity of S and G2/M phase measurements to variations in
the fitting model 57
References 58
4 Determination of the cell proliferative
fraction in human tumours 59
Rosella Silvestrini, Maria Grazia Daidone, and Aurora Costa
1 Introduction 59
2 Methods for procurement and preparation of tumour
specimens 60
Procurement of tumour tissue - 60
Preparation of specimens from solid tumours 60
Treatment of cell aspirates 61
3 Proliferation indices 62
Incorporation of nucleic acid precursors 62
Other measurements of cell proliferation 69
Methodological controls 73
Relationship between the different proliferation, indices 73
4 Predictive value of cell proliferative fraction measurements 74
Prognosis 75
Response to treatment 76
Acknowledgements 76
References 77
5 Chromosome analysis in cell culture 79
Anwar N Mohamed and Sandra R Wolman
1 Introduction 79
2 Blood and bone marrow cultures 80
Blood 80
Bone marrow optimal culture conditions 81
Methotrexate synchronization 81
3 Solid tissue cultures 85
Culture conditions of solid tumours 85
4 Staining and banding methods 88
Trypsin-Giemsa staining (G-banding) 89
Quinacrine staining (Q-banding) 89
Reverse (R-banding) 90
Constitutive heterochromatin banding (C-banding) 91
xi
Contents
} 5 Nomenclature and interpretation 92
Numerical and structural aberrations 92
Tumour cell populations 93
6 In situ hybridization and interphase cytogenetics 93
References 96
6 Assessment of DNA damage in
mammalian cells by DNA filter elution
methodology 97
Richard Bertrand and Yves Pommier
1 Introduction 97
2 DNA lesions measured by filter elution assays 97
3 Basic principles of the DNA filter elution methodology 98
4 Overview of the DNA filter elution procedures 100
Equipment 100
DNA labelling and preparation of experimental cell cultures 101
Calibration and internal standard cells 101
Cell lysis, DNA elution procedures, and treatment of filters 103
5 Specific conditions for each of the DNA filter elution
assays 104
Filters and solutions 104
Single- and double-strand breaks 106
Protein-associated strand breaks and protein-free breaks 106
DNA-protein crosslinks 106
Interstrand DNA crosslinks 107
Alkali-labile sites 107
Apoptosis-associated DNA fragmentation 107
Preparation of isolated nuclei for filter elution assays 108
Reconstituted cell-free system with filter elution assays 108
6 Computations 109
SSB (single-strand breaks) 109
DSB (double-strand breaks) 111
DPC (DNA-protein crosslinks) 112
ISC (interstrand crosslinks) 113
Apoptosis-associated DNA fragmentation 113
7 Conclusions and perspectives 114
Acknowledgements 115
References 116
xii
Contents
7 ^ Morphological and biochemical criteria of
Mpoptosis 119
Sumant Ramachandra and George P Studzinski
1 Introduction 119
2 Distinction of apoptosis from other forms of cell death 122
3 Morphological changes in apoptosis 122
Identification by light microscopy of cells undergoing apoptosis 125
Identification by electron microscopy of cells undergoing apoptosis 128
4 Histochemical detection of apoptosis 129
5 Biochemical detection of specific DNA damage for
demonstration of apoptosis 131
Determination of the increased mitochondrial to nuclear DNA
ratio for detection and quantitation of apoptosis 136
6 Assessment of nuclear proteolytic activity in cells
undergoing apoptosis 139
7 Concluding remarks 140
Acknowledgements 140
References 140
8 Analysis of cell death by flow cytometry 143
Zbigniew Darzynkiewicz, Xun Li, Jianping Gong,
Shinsuke Hara, and Frank Traganos
1 Introduction 143
2 Modes of cell death 143
Apoptosis 144
Necrosis 145
Atypical apoptosis: delayed reproductive, or mitotic, cell death 145
3 Features discriminating dead cells 146
Structural and functional integrity of the plasma membrane 146
Cell organelles 147
Specific features of apoptotic cells 147
4 Light scattering properties of dying cells 148
5 Removal of dead cells by incubation with DNase I and
trypsin 148
6 Exclusion of PI combined with hydrolysis of fluorescein
diacetate 150
7 Uptake of Rh l23 combined with exclusion of PI 151
8 Exclusion of PI followed by counterstaining with HO342 153
xiii
Contents
) 9 Uptake of HO342 combined with exclusion of PI 155
10 Extraction of the degraded DNA from apoptotic cells 156
11 Denaturability of DNA in situ 158
12 Labelling of DNA strand breaks in apoptotic cells 162
DNA strand labelling with biotinylated dUTP 162
DNA strand break labelling with digoxygenin-conjugated dUTP 163
13 Which method to choose? 164
Acknowledgements 166
References 166
9 Assays of cell growth and cytotoxicity 169
Philip Skehan
1 Introduction 169
2 Growth and cytotoxicity assays 169
3 Experimental design 170
4 Cell cultures 171
Seeding density 171
Dissociation and recovery 171
Drug solubilization 171
Assay duration 171
Control wells on every plate 172
5 Dye-binding assays 172
Optimizing and validating dye-binding assays 172
Protein and biomass stains 173
6 Metabolic impairment assays 179
Neutral Red viability assay 179
MTT cell viability assay 180
7 Membrane integrity assays 181
Fluorescein diacetate 181
8 Electrical conductivity assays 183
9 Survivorship assays 184
Adherent versus non-adherent cells 185
Long-term recovery (LTR) assay 185
Colony-forming efficiency on tissue culture plastic 187
Colony-forming efficiency in soft agar 188
References 190
xiv
Contents
10 Cell synchronization as a basis for
- investigating control of proliferation in
mammalian cells 193
Gary S Stein, Janet L Stein, Jane B Lian, T J Last,
Thomas Owen, and Laura McCabe
1 Introduction 193
2 Synchronization of continuously dividing cells by
thymidine 193
General considerations 193
Rationale for steps in thymidine synchronization protocol 197
Modification of thymidine synchronization for specialized
applications 198
Precautions that should be exercised during thymidine
synchronization 198
3 Synchronization of continuously dividing cells by
mitotic selective detachment 200
General considerations 200
Modification of mitotic selective detachment protocols 200
4 Induction of synchronous proliferative activity 202
References 203
11 Growth and activation of human
leukaemic cells in vitro and their growth
in the SCID mouse model 205
Sophie Visonneau, Alessandra Cesano, and Daniela Santoli
1 Introduction 205
2 Procedures for growing human leukaemic cells in
suspension culture 206
3 Clonal growth of leukaemic cells in semi-solid medium 208
Choice of the semi-solid medium 209
Preparation of the agar stock 209
Preparation of the methylcellulose stock 211
Scoring of colonies in semi-solid medium 212
Staining of colony-forming cells 212
4 Use of the SCID mouse model for growing human
leukaemias 212
The SCID mouse 212
Immunosuppressive treatments of SCID mice 214
Transfer of leukaemic cells i p 215
Intravenous (i v ) injection of leukaemic cells 215
xv
Contents
Success of human leukaemic cell engraftment
Appearance of symptoms
Autopsy and histopathology
Recovery of human leukaemic cells from mouse tissues
5 Human lymphocyte activation in culture
Cell-mediated cytotoxicity
Expression of intracytoplasmic azurophilic granules and lytic
proteins
E/T conjugate formation
Direct lysis of target cells
Role of Ca2+ in the killing process
Exocytosis of cytotoxic granules
Proliferative responses
Lymphokine production
Acknowledgements
References
12 Senescence and immortalization of
hujnan cells 229
Karen Hubbard and Harvey L Ozer
1 Introduction 229
2 Cellular senescence 230
3 SV40-transformation and crisis 235
4 SV40-immortalized cell lines 238
5 Conditional SV40 transformants 241
6 Other approaches to immortalization of human cells 243
7 Summary 244
References 244
Appendix Changes identified in senescing mammalian cells 245
13 Analysis of the effects of purified growth
factors on normal human fibroblasts 251
Paul D Phillips and Vincent J Cristofalo
1 Introduction 251
xvi
Contents
2 Studies with growth factors 253
) Materials 253
Trypsinizing and harvesting the cells 255
A classification of growth factors for WI-38 cells 257
References 262
Al Addresses of suppliers 265
Index 267
xvu
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| indexdate | 2024-07-09T17:51:44Z |
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| isbn | 0199635692 0199635684 |
| language | English |
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| series | The practical approach series |
| series2 | The practical approach series |
| spelling | Cell growth and apoptosis a practical approach ed. by G. P. Studzinski Oxford IRL Press 1995 XXVI, 269 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier The practical approach series 159 This Practical Approach volume is unique in describing the key concepts of cell growth and cell death, particularly by apoptosis, and in providing detailed methods to study these processes. The chapters are written by experts in the field and cover a wide range of important subject areas including cell-cycle traverse, chromosomal abnormalities detected at cell division, DNA damage, and cell death or senescence. The establishment of cell lines and optimization of cell proliferation by growth factors are also described. The result is an invaluable laboratory manual for studies of growth and its abnormalities in mammalian cells. Cell Growth and Apoptosis: A Practical Approach provides state-of-the-art protocols for studies of normal and abnormal cell growth, including cancer and other cell-proliferation disorders, together with a critical appraisal of the various methodologies in use. Its coverage of human cells will make it particularly useful for health professionals. Cell Growth cabt Apoptosis cabt Cell Division cabt Celdood gtt Celgroei gtt Apoptosis Apoptosis Research Methodology Cell Division Cells Growth Research Methodology Cytological Techniques Apoptosis (DE-588)4291096-1 gnd rswk-swf Methode (DE-588)4038971-6 gnd rswk-swf Zellwachstum (DE-588)4190674-3 gnd rswk-swf Zellwachstum (DE-588)4190674-3 s Apoptosis (DE-588)4291096-1 s DE-604 Methode (DE-588)4038971-6 s Studzinski, George P. Sonstige oth The practical approach series 159 (DE-604)BV007921321 159 HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=006920791&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Cell growth and apoptosis a practical approach The practical approach series Cell Growth cabt Apoptosis cabt Cell Division cabt Celdood gtt Celgroei gtt Apoptosis Apoptosis Research Methodology Cell Division Cells Growth Research Methodology Cytological Techniques Apoptosis (DE-588)4291096-1 gnd Methode (DE-588)4038971-6 gnd Zellwachstum (DE-588)4190674-3 gnd |
| subject_GND | (DE-588)4291096-1 (DE-588)4038971-6 (DE-588)4190674-3 |
| title | Cell growth and apoptosis a practical approach |
| title_auth | Cell growth and apoptosis a practical approach |
| title_exact_search | Cell growth and apoptosis a practical approach |
| title_full | Cell growth and apoptosis a practical approach ed. by G. P. Studzinski |
| title_fullStr | Cell growth and apoptosis a practical approach ed. by G. P. Studzinski |
| title_full_unstemmed | Cell growth and apoptosis a practical approach ed. by G. P. Studzinski |
| title_short | Cell growth and apoptosis |
| title_sort | cell growth and apoptosis a practical approach |
| title_sub | a practical approach |
| topic | Cell Growth cabt Apoptosis cabt Cell Division cabt Celdood gtt Celgroei gtt Apoptosis Apoptosis Research Methodology Cell Division Cells Growth Research Methodology Cytological Techniques Apoptosis (DE-588)4291096-1 gnd Methode (DE-588)4038971-6 gnd Zellwachstum (DE-588)4190674-3 gnd |
| topic_facet | Cell Growth Apoptosis Cell Division Celdood Celgroei Apoptosis Research Methodology Cells Growth Research Methodology Cytological Techniques Methode Zellwachstum |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=006920791&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV007921321 |
| work_keys_str_mv | AT studzinskigeorgep cellgrowthandapoptosisapracticalapproach |