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Tables of Contents for Aquatic Invertebrate Cell Culture
Chapter/Section Title
Page #
Page Count
Preface
xiii
Introduction
1
6
C. Mothersill
B. Austin
References
4
3
Part I Fundamental techniques
7
54
Basic methods and media
9
6
C. Mothersill
A. L. Mulford
B. Austin
Animals
9
1
Equipment
9
1
Procedures
10
3
Good aseptic technique
10
1
Hygiene of animals
10
1
Appropriate disinfectant technique for the whole animal
10
1
Appropriate dissection techniques
10
1
Need of speed for processing tissues
10
1
Which incubation conditions -- with or without carbon dioxide?
11
1
Nature of vessels -- plastic or glass, coated or uncoated?
11
1
Area of culture vessels
11
1
Use of antibiotics
11
1
Explants or isolated cells?
11
1
Observation of cultures
12
1
Subcultures
12
1
Use of viability tests
12
1
Characterization of the cells
12
1
Culture media
13
1
Which medium
13
1
Which supplements?
13
1
Medium-change regime
13
1
Conclusions
14
1
References
14
1
Tissue choices for aquatic invertebrate tissue culture
15
10
A. Evenden
Introduction
15
3
Choosing an appropriate animal
15
1
Availability of animals
16
1
Animal suitability
17
1
Tissue choices
18
6
Developmental status
18
3
Methodological factors
21
1
Application choices
22
1
A way forward
23
1
References
24
1
Characterization
25
36
M. Lyons-Alcantara
Introduction
25
3
Morphology
28
1
Size
29
1
Ultrastructure
30
4
Epithelial tissue
30
2
Fibroblasts
32
2
Cell-growth kinetics
34
2
Cell counting at different time intervals
34
1
Mitotic index
34
1
DNA Incorporation methods
34
1
AgNOR
35
1
Flow cytometry
35
1
Telomerase activity
36
1
Immunohistochemical methods
36
1
Enzyme activity
36
1
Antigenicity
37
7
Structural markers
37
5
Proliferation markers
42
2
Differentiation
44
1
Histochemistry
44
1
DNA Fingerprinting
45
1
Isoenzyme analysis
45
1
Chromosome content
46
1
DNA content
46
1
RNA and protein
47
1
Expression of a sponge gene as a marker for the origin of the cells
47
1
Contamination with unicellular eukaryotes
48
1
References
49
12
Part II Experiences with aquatic invertebrates
61
208
Cell cultures from crustaceans: Shrimps, crabs and crayfish
63
72
A. L. Mulford
A. J. Villena
Introduction
63
1
The crustaceans
64
3
Previous experiences of tissue culture
67
14
Short-term cultures
68
1
Long-term cultures
69
12
Physiological data useful to define culture media for crustacean cells
81
12
Methods
93
24
Anaesthesia and decontamination procedures
95
1
Initiation of cultures
96
10
Attachment factors
106
1
Culture media
107
3
Culture conditions
110
1
Culture identification
110
7
Conclusions
117
1
References
118
17
Interaction between crustacean ectoparasites and their hosts: A tissue-culture perspective
135
30
D. T. Nolan
S. C. Johnson
Introduction
135
1
Important crustacean ectoparasites
136
8
Caligid fish lice
136
1
Branchiuran fish lice
137
1
Other crustacean ectoparasites of fish
137
1
Parasite-induced stress effects and disease susceptibility
137
2
Bottlenecks in studying the host-parasite interaction in greater depth and potential new directions
139
2
Challenges in attempting cell culture from crustacean ectoparasitic lice
141
3
Cell-culture trials with Argulus japonicus
144
9
Experimental animals and handling
144
1
Media composition and supplements
144
1
Tissue preparation and culture conditions
145
3
Evaluation of culture results
148
1
Characterization requirements for cells
148
5
Application of cell culture for control of these and other types of parasites
153
1
Conclusions
154
1
Acknowledgements
155
1
References
155
10
Culture of molluscan cells
165
18
M. F. Mulcahy
Introduction
165
11
Target species
165
1
Target cells
166
3
Depuration, decontamination and disinfection
169
1
Cell-isolation protocols
170
1
Cell-culture conditions
171
2
Methods to measure the functional activity of cells
173
1
Characterization of cultured cells
173
1
Approaches to the development of continuous cell lines
174
1
Cryopreservation
175
1
Conclusion
176
1
References
176
7
Molluscan diseases: A tissue-culture perspective
183
22
R. A. Elston
Introduction
183
1
Viral diseases and isolation of viruses from molluscs
184
8
Viral diseases of molluscs
184
2
Culture of molluscan viruses
186
3
Dissemination of neoplasia of molluscs: A viral aetiology?
189
3
Intracellular bacterial diseases of molluscs
192
2
Intracellular protozoan diseases of molluscs
194
1
Efforts to establish molluscan cell and tissue cultures
194
3
Primary culture of molluscan cells and tissues
194
2
Establishment of the Bge cell line
196
1
Need for the establishment of molluscan cell and tissue cultures
197
1
References
198
7
Primary cell culture from a sponge: Primmorphs
205
20
W. E. G. Muller
M. R. Custodio
Introduction
205
1
Establishment of primmorphs
206
5
Sponge
206
1
Media
206
1
Dissociation of cells and formation of primmorphs
207
2
Histology
209
1
Subcultures of primmorphs
210
1
Long-term cultivation of primmorphs in the aquarium
210
1
Characterization of primmorphs
211
2
Level of telomerase activity in cells depending on the culture conditions
211
1
Immunocytochemical detection of BrdU incorporation in cells of primmorphs
211
1
Presence of bacteria in the primmorphs
212
1
Expression of a sponge gene as a marker for the origin of the cells
212
1
Application of primmorphs
213
1
Bioindicator
213
1
Production of bioactive compounds
214
1
Cultivation of primmorphs under pressure
214
4
Effect of ethylene
214
2
Cloning of ethylene-responsive proteins from Suberites domuncula
216
1
Levels of expression of sponge ethylene-responsive protein
216
2
Discussion of the system
218
1
Acknowledgements
219
1
Disclosure statement
219
1
References
220
5
Urochordate cell cultures: From in vivo to in vitro approaches
225
20
B. Rinkevich
C. Rabinowitz
Tunicates: importance for cell-culture applications
225
1
Four major tissue sources for in vitro studies
226
4
Naturally occurring cell suspensions
226
1
Organ cultures
227
2
Dissociated tissues/organs
229
1
Embryo-derived cell cultures
229
1
Establishment of cell cultures
230
1
Botryllid ascidians: Cell cultures; recent advances, old problems
231
3
Blood cells
231
1
Ampullae regeneration
232
1
Epithelial cell cultures
233
1
In vitro applications
234
4
Applications for environmental studies
236
2
Closing remarks
238
2
Acknowledgements
240
1
References
240
5
Echinodermata
245
24
C. Moss
C. D. Bavington
J. D. McKenzie
The echinoderms
245
1
Echinoderm cells and tissues
246
5
Tissue layout and cell culture
246
3
Reproductive and other proliferative tissues
249
2
Studies on echinoderm cells in vitro and the development of cell-culture techniques
251
11
The development of methodology towards long-term echinoderm cultures
254
8
Applications for echinoderm cell cultures
262
2
Production of metabolites
263
1
Use of symbionts
263
1
Models for aquaculture and environmental monitoring
263
1
Pest species
264
1
Summary
264
1
References
264
5
Part III Uses for cell cultures
269
132
Application of cell culture for coral-reef ecology
271
22
Z. Billinghurst
M. S. A. Ammar
W. E. G. Muller
Introduction
271
3
Cnidarians
271
3
Cell culture
274
3
Primmorph system
275
2
Applications of the primmorph system
277
10
Diseases in corals
278
1
Coral bleaching
279
3
Molecular probes for detecting environmental stress
282
1
Case study: Monitoring the effect of light using molecular probes
283
1
Primmorphs from Dendronephthya klunzingeri as a model to determine the effect of UVB and visible light
284
3
Acknowledgements
287
1
Disclosure statement
287
1
References
287
6
Culture of bryozoans and barnacles: Application of larval cell culture to biofouling studies
293
30
K. Okano
E. Hunter
Introduction
293
2
Cells from Bryozoan larvae
295
11
Overview
295
3
Techniques and methodology
298
4
Bryozoan larval ciliated epithelium
302
1
Internal cells from bryozoan larvae
303
2
Modulation of ciliary behaviour in identified ciliated cells
305
1
Cells from barnacle (cirripedia) larvae
306
7
Overview
306
3
Techniques and methodology
309
2
Physiological characterization of cement secretion using isolated organs and cultured cells
311
2
Future applications
313
3
Application of larval cell-culture techniques in biofouling studies
313
1
Larval cells as cement factories
314
1
Larval cells and pharmaceuticals
315
1
Conclusion
316
1
Acknowledgements
316
1
References
316
7
Development of sponge cell cultures for biomedical application
323
14
S. A. Pomponi
R. Willoughby
Importance of sponge cell culture
323
1
Sponge cell cultures
324
1
Cell culture versus tissue culture
324
1
Establishment of cell cultures
325
2
Selection of appropriate species for in vitro research
325
1
Identification of target cell types
326
1
Selection of appropriate methods for cell dissociation
326
1
Cryopreservation: pros and cons
327
1
Development of defined culture media
327
3
Basal media
329
1
Antibiotics
329
1
Growth factors
329
1
Selection of culture vessel
330
2
Attached versus suspension cultures
330
2
Cell-type verification and assessment of functional state
332
1
Chemical analyses of cultured cells
333
1
Biosynthesis of radiolabelled precursors
333
1
Current status and future directions
333
1
Acknowledgements
334
1
References
334
3
Applications of invertebrate cell culture in studies of biomarkers and ecotoxicology
337
24
D. Sheehan
Introduction
337
5
Chemical threat posed by xenobiotics
337
1
Biological xenobiotic defence mechanisms
338
1
Enzymatic detoxification
339
3
Detoxification systems of invertebrates as bioindicators of environmental pollution
342
1
Invertebrates as sentinel species
342
1
Potential of invertebrate species for in vitro toxicology
343
1
Primary cell cultures
343
4
Uses of cell lines in toxicology
347
3
Cytotoxicity
347
1
Ultrastructural effects
348
1
Formation of protein adducts
348
1
Genotoxicity
349
1
Non-mammalian tissue culture
350
1
Potential of invertebrate tissue culture in toxicology
351
1
Future perspectives
351
1
References
352
9
The growth and pathogenicity of bacterial and viral pathogens in aquatic invertebrate tissue cultures
361
8
B. Austin
Introduction
361
4
The availability of cell cultures
362
1
The growth of viral pathogens
362
1
The growth of bacterial pathogens
363
1
Parasites
364
1
Measuring cellular damage
364
1
Conclusion
365
1
References
365
4
Validation of in vitro techniques
369
28
B. Quinn
C. Mothersill
In vitro techniques
369
5
Validation
370
3
Validation by in vitro/in vivo comparison
373
1
Legislation and regulations
374
1
Tier system for use of validating in vitro techniques
374
5
QSAR
375
1
In vitro exposure
376
1
In vivo exposure
376
1
In vitro/in vivo validation
377
2
Validated in vitro systems
379
1
Mutagenicity
379
1
Skin and eye irritancy
379
1
Toxicity studies
380
2
Pathogenicity studies
382
1
Validation by information systems and databases
383
2
Databases
384
1
The MEIC programme
385
1
Conclusion
385
2
References
387
10
Conclusions
397
4
C. Mothersill
B. Austin
Index
401