Escolar Documentos
Profissional Documentos
Cultura Documentos
culture
a practical approach
Edited by
RI Freshney
Cancer Research Campaign, Department of Oncology,
University of Glasgow, 1 Horselethill Road,
Glasgow, G12 9LX, UK
FACHBEREICH B I O L O G I E (10)
der Technischen Hochschule Darmstadt
- Brbliothek D - 6 1 0 0 D a r m s t a d t / B. R. D.
Schnlttsoorinstrafie
OIRL PRESS
Oxford Washington DC
Contents
ABBREVIATIONS
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vn
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Introduction
General Methods and Culture Parameters
Cell quantification
Equipment and reagents
Practical considerations
Growth kinetics
Medium and nutrients
pH
Oxygen
Types of culture systems
Monolayer Culture
Introduction
Cell attachment
Scaling-up: step 1, roller bottle
Scaling-up: step 2, roller bottle modifications
Scaling-up: step 3, large capacity stationary cultures
Scaling-up: step 4, fermenter systems
Summary
Suspension Culture
Adaptation to suspension culture
Static suspension culture
Small-scale suspension culture
Scaling-up factors
Stirred fermenters
Continuous flow culture
Airlift fermenter
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Encapsulated cells
References
Appendix
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via
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Introduction
Cell Line Banking
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Introduction
Cell Separation Methods
The Centrifugal Elutriator
Elutriator rotor
Elutriation chamber
Peristaltic pump and additional apparatus
Complete elutriation system
Technical Protocol
Sterilisation of the apparatus
Flow rate calibration
Sample loading
Elutriation of discrete cell populations
Collection and Interpretation of Data
Heterogeneous cell populations (assessment of cell fraction
purity)
Established cell lines (recovery and viability)
Future Development
Acknowledgements
References
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FLOW CYTOMETRY
L.Morasca and E.Erba
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Introduction
Design of Equipment
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ORGAN CULTURE
I.Lasnitzki
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Introduction
Survey of Organ Culture Techniques
Clotted plasma substrate
Agar substrate
Raft methods
Grid method
Intermittent exposure to medium and gas phase
Watchglass Technique
Preparation of embryo extract
Preparation of chicken plasma
Explantation
The Maximow Single Slide Technique
Materials
Setting up of cultures
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Renewal of medium
Agar Gel Techniques
Embryological watchglass technique
Embryo Culture for Teratological Studies
Explantation procedure
Gas mixture
Medium
Protocol
Grid Technique
Materials
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Explantation procedures
Medium
Explantation
Incubation and perfusion
Renewal of medium
Long-term Cultivation of Human Adult Tissues
Explantation and cultivation
Medium
Incubation
Change of medium
Preparation of Organ Cultures for Light Microscopy
Materials
Fixation dehydration and embedding
Staining methods
Autoradiography
Labelling
Fixation
Dehydration and embedding
Dipping procedure
Guide to Quantitation of Results
Advantages and Limitations of Organ Culture
References
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Introduction
Background
Specific Techniques
Culture methods
Drug Concentrations
Duration of Drug Exposure
Recovery Period
End-points
Cytotoxicity, viability and survival
- ...
Cytotoxicity and viability
Survival (reproductive integrity)
Assay Comparisons
Technical Protocols
Drugs and drug solutions
Drug incubation
Assay by survival and proliferative capacity
Cytotoxicity assays
Interpretation of Results
Relationship between cell number and cytotoxicity index
Dose-response curves
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xi
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APPENDICES
I
II
General Reagents
Suppliers of Specialist Items
INDEX
XII
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