Culture of Animal Cells E-Book

R. Ian Freshney

• Herausgeber: John Wiley & Sons
• Kategorie: Wissenschaft und neue Technologien
• Sprache: Englisch

Seitenzahl: 2099

Veröffentlichungsjahr: 2015

Beschreibung

Since the publication of the sixth edition of this benchmark text, numerous advances in the field have been made - particularly in stem cells, 3D culture, scale-up, STR profiling, and culture of specialized cells. Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications, Seventh Edition is the updated version of this benchmark text, addressing these recent developments in the field as well as the basic skills and protocols. This eagerly awaited edition reviews the increasing diversity of the applications of cell culture and the proliferation of specialized techniques, and provides an introduction to new subtopics in mini-reviews. New features also include a new chapter on cell line authentication with a review of the major issues and appropriate protocols including DNA profiling and barcoding, as well as some new specialized protocols. Because of the continuing expansion of cell culture, and to keep the bulk of the book to a reasonable size, some specialized protocols are presented as supplementary material online. Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications, Seventh Edition provides the most accessible and comprehensive introduction available to the culture and experimental manipulation of animal cells. This text is an indispensable resource for those in or entering the field, including academic research scientists, clinical and biopharmaceutical researchers, undergraduate and graduate students, cell and molecular biology and genetics lab managers, trainees and technicians.

Leseprobe

CULTURE OF ANIMAL CELLS

A MANUAL OF BASIC TECHNIQUE AND SPECIALIZED APPLICATIONS

Seventh Edition

R. Ian Freshney

Institute of Cancer Sciences University of Glasgow

Reviewing Editors

Amanda Capes-Davis

Children’s Medical Research Institute University of Sydney, Westmead, Australia

Carl Gregory

Institute for Regenerative Medicine Texas A&M Health Science Center, Temple, TX, USA

Stefan Przyborski

School of Biological & Biomedical Science Durham University, Durham, UK

Copyright © 2016 by John Wiley & Sons, Inc. All rights reserved

Published by John Wiley & Sons, Inc., Hoboken, New Jersey Published simultaneously in Canada

No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, scanning, or otherwise, except as permitted under Section 107 or 108 of the 1976 United States Copyright Act, without either the prior written permission of the Publisher, or authorization through payment of the appropriate per-copy fee to the Copyright Clearance Center, Inc., 222 Rosewood Drive, Danvers, MA 01923, [phone], fax [phone], or on the web at www.copyright.com. Requests to the Publisher for permission should be addressed to the Permissions Department, John Wiley & Sons, Inc., 111 River Street, Hoboken, NJ 07030, [phone], fax [phone], or online at http://www.wiley.com/go/permission.

Limit of Liability/Disclaimer of Warranty: While the publisher and author have used their best efforts in preparing this book, they make no representations or warranties with respect to the accuracy or completeness of the contents of this book and specifically disclaim any implied warranties of merchantability or fitness for a particular purpose. No warranty may be created or extended by sales representatives or written sales materials. The advice and strategies contained herein may not be suitable for your situation. You should consult with a professional where appropriate. Neither the publisher nor author shall be liable for any loss of profit or any other commercial damages, including but not limited to special, incidental, consequential, or other damages.

For general information on our other products and services or for technical support, please contact our Customer Care Department within the United States at [phone], outside the United States at [phone] or fax [phone].

Wiley also publishes its books in a variety of electronic formats. Some content that appears in print may not be available in electronic formats. For more information about Wiley products, visit our website at www.wiley.com.

Library of Congress Cataloging-in-Publication Data:

Freshney, R. Ian.    Culture of animal cells : a manual of basic technique and specialized applications, / R. Ian Freshney. – 7th ed.       p. cm.    Includes index.    ISBN 978-1-118-87365-6 (cloth) 1. Tissue culture–Laboratory manuals. 2. Cell culture–Laboratory manuals. I. Title.    QH585.2.F74 2016    571.6381–dc22

2016007042

This book is dedicated to all of the many friends and colleagues whose help and advice over the years has enabled me to extend the scope of this book beyond my own limited experience.

CONTENTS

List of Color Plates

List of Protocols

List of Minireviews

Preface and Acknowledgments

Abbreviations

About the Companion Website

CHAPTER 1 Introduction

1.1 HISTORICAL BACKGROUND

1.2 ADVANTAGES OF TISSUE CULTURE

1.3 LIMITATIONS

1.4 MAJOR DIFFERENCES IN VITRO

1.5 TYPES OF TISSUE CULTURE

REFERENCES

CHAPTER 2 Biology of Cultured Cells

2.1 THE CULTURE ENVIRONMENT

2.2 CELL ADHESION

2.3 CELL PROLIFERATION

2.4 DIFFERENTIATION

2.5 CELL SIGNALING

2.6 ENERGY METABOLISM

2.7 ORIGIN OF CULTURED CELLS

2.8 DEFINITIONS

REFERENCES

CHAPTER 3 Laboratory Design and Layout

3.1 PLANNING, FURNISHING, AND SERVICES

3.2 DESIGN AND LAYOUT

3.3 DISASTER MANAGEMENT

REFERENCES

CHAPTER 4 Equipment and Materials

4.1 REQUIREMENTS OF A TISSUE CULTURE LABORATORY

4.2 STERILE WORKING AREA

4.3 INCUBATION AND CULTURE

4.4 PREPARATION AND STERILIZATION

4.5 STORAGE

4.6 SUPPLEMENTARY LABORATORY EQUIPMENT

4.7 SPECIALIZED EQUIPMENT

REFERENCES

CHAPTER 5 Aseptic Technique

5.1 OBJECTIVES OF ASEPTIC TECHNIQUE

5.2 ELEMENTS OF ASEPTIC ENVIRONMENT

5.3 STERILE HANDLING

5.4 STANDARD PROCEDURE

5.5 APPARATUS AND EQUIPMENT

REFERENCES

CHAPTER 6 Safety, Bioethics, and Validation

6.1 LABORATORY SAFETY

6.2 RISK ASSESSMENT

6.3 STANDARD OPERATING PROCEDURES

6.4 SAFETY REGULATIONS

6.5 GENERAL SAFETY

6.6 FIRE

6.7 IONIZING RADIATION

6.8 BIOHAZARDS

6.9 BIOETHICS

6.10 QUALITY ASSURANCE

REFERENCES

CHAPTER 7 Culture Vessels and Substrates

7.1 THE SUBSTRATE

7.2 TREATED SURFACES

7.3 CHOICE OF CULTURE VESSEL

7.4 SPECIALIZED SYSTEMS

REFERENCES

CHAPTER 8 Defined Media and Supplements

8.1 DEVELOPMENT OF MEDIA

8.2 PHYSICOCHEMICAL PROPERTIES

8.3 BALANCED SALT SOLUTIONS

8.4 COMPLETE MEDIA

8.5 SERUM

8.6 SELECTION OF MEDIUM AND SERUM

8.7 OTHER SUPPLEMENTS

8.8 STORAGE

REFERENCES

CHAPTER 9 Serum-Free Media

9.1 DISADVANTAGES OF SERUM

9.2 ADVANTAGES OF SERUM-FREE MEDIA

9.3 DISADVANTAGES OF SERUM-FREE MEDIA

9.4 REPLACEMENT OF SERUM

9.5 DEVELOPMENT OF SERUM-FREE MEDIUM

9.6 SELECTION OF SERUM-FREE MEDIUM

9.7 PREPARATION OF SERUM-FREE MEDIUM

9.8 ANIMAL PROTEIN−FREE MEDIA

9.9 CONCLUSIONS

REFERENCES

CHAPTER 10 Preparation and Sterilization

10.1 PREPARATION OF REAGENTS AND MATERIALS

10.2 STERILIZATION OF APPARATUS AND LIQUIDS

10.3 APPARATUS

10.4 REAGENTS AND MEDIA

10.5 STERILIZATION OF MEDIA

10.6 CONTROL, TESTING, AND STORAGE OF MEDIA

REFERENCES

CHAPTER 11 Primary Culture

11.1 INITIATION OF A PRIMARY CELL CULTURE

11.2 ISOLATION OF THE TISSUE

11.3 TYPES OF PRIMARY CULTURE

REFERENCES

CHAPTER 12 Subculture and Cell Lines

12.1 TERMINOLOGY

12.2 SUBCULTURE AND PROPAGATION

12.3 CHOOSING A CELL LINE

12.4 ROUTINE MAINTENANCE

12.5 SUBCULTURE

REFERENCES

CHAPTER 13 Authentication and Validation

13.1 AUTHENTICATION OF CELL LINES

13.2 VALIDATION

13.3 QUALITY ASSURANCE

REFERENCES

CHAPTER 14 Microbial Contamination

14.1 SOURCES OF CONTAMINATION

14.2 TYPES OF MICROBIAL CONTAMINATION

14.3 MONITORING FOR CONTAMINATION

14.4 DISPOSAL OF CONTAMINATED CULTURES

14.5 ERADICATION OF CONTAMINATION

14.6 CROSS-CONTAMINATION

14.7 CONCLUSIONS

REFERENCES

CHAPTER 15 Cryopreservation and Banking

15.1 RATIONALE FOR FREEZING

15.2 CONSIDERATIONS BEFORE CRYOPRESERVATION

15.3 PRINCIPLES OF CRYOPRESERVATION

15.4 DESIGN AND CONTROL OF FREEZER STOCKS

15.5 CELL BANKS

15.6 TRANSPORTING CELLS

REFERENCES

CHAPTER 16 Cloning and Selection

16.1 CELL CLONING

16.3 SUSPENSION CLONING

16.4 ISOLATION OF CLONES

16.5 REPLICA PLATING

16.6 SELECTIVE MEDIA AND INHIBITORS

16.7 INTERACTION WITH SUBSTRATE

REFERENCES

CHAPTER 17 Cell Sorting

17.1 CELL DENSITY AND ISOPYKNIC SEDIMENTATION

17.2 CELL SIZE AND SEDIMENTATION VELOCITY

17.3 ANTIBODY-BASED TECHNIQUES

17.4 BEGINNER’S APPROACH TO CELL SEPARATION

REFERENCES

CHAPTER 18 Cell Line Characterization

18.1 THE NEED FOR CHARACTERIZATION

18.2 GENOTYPIC CHARACTERIZATION

18.3 PHENOTYPIC CHARACTERIZATION

18.4 CELL MORPHOLOGY

18.5 ANTIGENIC MARKERS

18.6 TIME-LAPSE RECORDING

18.7 CELL CYCLE

REFERENCES

CHAPTER 19 Differentiation

19.1 EXPRESSION OF THE IN VIVO PHENOTYPE

19.2 STAGES OF DIFFERENTIATION

19.3 PROLIFERATION AND DIFFERENTIATION

19.4 COMMITMENT AND LINEAGE

19.5 STEM CELL PLASTICITY

19.6 MARKERS OF DIFFERENTIATION

19.7 INDUCTION OF DIFFERENTIATION

19.8 DIFFERENTIATION AND MALIGNANCY

19.9 PRACTICAL ASPECTS

REFERENCES

CHAPTER 20 Three-Dimensional Culture

20.1 CELL INTERACTION AND PHENOTYPIC EXPRESSION

20.2 ORGAN CULTURE

20.3 HISTOTYPIC CULTURE

20.4 ORGANOTYPIC CULTURE

20.5 IMAGING CELLS IN 3D CONSTRUCTS

REFERENCES

CHAPTER 21 Scale-up and Automation

21.1 SCALE-UP IN SUSPENSION

21.2 SCALE-UP IN MONOLAYER

21.3 PROCESS CONTROL

21.4 LARGE-SCALE PROCESSES

21.5 AUTOMATION

REFERENCES

CHAPTER 22 Senescence, Immortalization, and Transformation

22.1 ROLE IN CELL LINE CHARACTERIZATION

22.2 GENETIC INSTABILITY AND HETEROGENEITY

22.3 IMMORTALIZATION

22.4 ABERRANT GROWTH CONTROL

22.5 TUMORIGENICITY

REFERENCES

CHAPTER 23 Quantitation

23.1 CELL COUNTING

23.2 CELL WEIGHT AND PACKED CELL VOLUME

23.3 DNA CONTENT

23.4 PROTEIN

23.5 RATES OF SYNTHESIS

23.6 PREPARATION OF SAMPLES FOR ENZYME ASSAY AND IMMUNOASSAY

23.7 CYTOMETRY

23.8 REPLICATE SAMPLING

23.9 CELL PROLIFERATION

23.10 PLATING EFFICIENCY

23.11 LABELING INDEX

23.12 AUTORADIOGRAPHY

23.13 CELL CYCLE TIME

23.14 CELL MIGRATION

REFERENCES

CHAPTER 24 Cytotoxicity

24.1 VIABILITY, TOXICITY, AND SURVIVAL

24.2 IN VITRO LIMITATIONS

24.3 NATURE OF THE ASSAY

24.4 APPLICATIONS OF CYTOTOXICITY ASSAYS

24.5 GENOTOXICITY

24.6 INFLAMMATION

REFERENCES

CHAPTER 25 Culture of Specific Cell Types

25.1 CELL CULTURE OF SPECIALIZED CELLS

25.2 EPITHELIAL CELLS

25.3 MESENCHYMAL CELLS

25.4 NEUROECTODERMAL CELLS

25.5 HEMATOPOIETIC CELLS

25.6 PRODUCTION OF MONOCLONAL ANTIBODIES

25.7 GONADS

25.8 CULTURE OF CELLS FROM POIKILOTHERMS

25.9 TUMOR CELL CULTURE

25.10 SAMPLING

25.11 DISAGGREGATION

25.12 PRIMARY CULTURE

25.13 SELECTIVE CULTURE OF TUMOR CELLS

25.14 DEVELOPMENT OF CELL LINES

25.15 CHARACTERIZATION OF TUMOR CELL CULTURES

25.16 SPECIFIC TUMOR TYPES

REFERENCES

CHAPTER 26 Stem Cells

26.1 EMBRYONIC STEM CELLS

26.2 GERM CELLS

26.3 EXTRAEMBRYONIC CELLS

26.4 HEMOPOIETIC STEM CELLS

26.5 APPICATION OF STEM CELLS TO REGENERATIVE MEDICINE

26.6 CANCER STEM CELLS

REFERENCES

CHAPTER 27 Training Programs

27.1 OBJECTIVES

27.2 PREPARATIVE AND MANIPULATIVE SKILLS

27.3 BASIC CELL CULTURE TECHNIQUES

27.4 ADVANCED EXERCISES

27.5 SPECIALIZED EXERCISES

REFERENCES

CHAPTER 28 Problem Solving

28.1 ABNORMAL APPEARANCE OF CELLS

28.2 SLOW CELL GROWTH

28.3 MEDIUM

28.4 SUBSTRATES AND CONTAINERS

28.5 MICROBIAL CONTAMINATION

28.6 CHEMICAL CONTAMINATION

28.7 PRIMARY CULTURE

28.8 DIFFERENTIATION

28.9 FEEDING

28.10 SUBCULTURE

28.11 CLONING

28.12 CROSS-CONTAMINATION AND MISIDENTIFICATION

28.13 CRYOPRESERVATION

28.14 CELL COUNTING

REFERENCE

CHAPTER 29 In Conclusion

Appendix I Calculations and preparation of reagents

REFERENCES

APPENDIX II Glossary

REFERENCE

Index

Links

Supplementary Images

EULA

List of Tables

Chapter 1

Table 1.1

Table 1.2

Table 1.3

Table 1.4

Table 1.5

Chapter 2

Table 2.1

Table 2.2

Chapter 3

Table 3.1

Table 3.2

Chapter 4

Table 4.1

Table 4.2

Table 4.3

Chapter 5

Table 5.1

Chapter 6

Table 6.1

Table 6.2

Table 6.3

Table 6.4

Table 6.5

Table 6.6

Table 6.7

Table 6.8

Table 6.9

Chapter 7

Table 7.1

Table 7.2

Chapter 8

Table 8.1

Table 8.2

Table 8.3

Table 8.4

Table 8.5

Table 8.6

Table 8.7

Chapter 9

Table 9.1

Table 9.2

Table 9.3

Table 9.4

Table 9.5

Chapter 10

Table 10.1

Table 10.2

Table 10.3

Table 10.4

Table 10.5

Table 10.6

Chapter 11

Table 11.1

Table 11.2

Table 11.3

Chapter 12

Table 12.1

Table 12.2

Table 12.3

Table 12.4

Table 12.5

Table 12.6

Table 12.7

Chapter 13

Table 13.1

Table 13.2

Table 13.3

Chapter 14

Table 14.1

Table 14.2

Table 14.3

Chapter 15