Culture of animal cells : a manual of basic technique and specialized applications / R. Ian Freshney ; reviewing editors, Amanda Capes-Davis, Carl Gregory, Stefan Przyborski.

Format:

Book

Contributor:

Capes-Davis, Amanda, editor.

Gregory, Carl, editor.

Przyborski, Stefan, editor.

Language:

English

Subjects (All):

Cell culture--Laboratory manuals.

Cell culture.

Tissue culture--Laboratory manuals.

Tissue culture.

Physical Description:

1 online resource (758 pages, 30 unnumbered pages of plates) : illustrations

Edition:

Seventh edition.

Place of Publication:

Hoboken, N.J.: Wiley Blackwell, c2016.

Hoboken, New Jersey : Wiley-Blackwell, 2016.

Summary:

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.

Contents:

Intro

CULTURE OF ANIMAL CELLS

Contents

List of Figures

List of Color Plates

List of Tables

List of Protocols

List of Minireviews

Preface and Acknlowledgments

Abbreviations

About the Companion Website

1. Introduction

1.1 Historical Background

1.2 Advantages of Tissue Culture

1.2.1 Control of the Environment

1.2.2 Characterization and Homogeneity of Sample

1.2.3 Economy, Scale, and Mechanization

1.2.4 In vitro Modeling of In vivo Conditions

1.3 Limitations

1.3.1 Expertise

1.3.2 Quantity

1.3.3 Dedifferentiation and Selection

1.3.4 Origin of Cells

1.3.5 Instability

1.4 Major Differences In Vitro

1.5 Types of Tissue Culture

References

2. Biology of Cultured Cells

2.1 The Culture Environment

2.2 Cell Adhesion

2.2.1 Cell Adhesion Molecules

2.2.2 Intercellular Junctions

2.2.3 Cytoskeleton

2.2.4 Extracellular Matrix

2.2.5 Cell Motility

2.3 Cell Proliferation

2.3.1 Cell Cycle

2.3.2 Control of Cell Proliferation

2.4 Differentiation

2.4.1 Induction and Maintenance of Differentiation

2.4.2 Plasticity of Differentiation and Dedifferentiation

2.5 Cell Signaling

2.6 Energy Metabolism

2.7 Origin of Cultured Cells

2.7.1 Initiation of the Culture

2.7.2 Evolution of Cell Lines

2.7.3 Senescence

2.7.4 Transformation and the Development of Continuous Cell Lines

2.8 Definitions

Supplier

3. Laboratory Design and Layout

3.1 Planning, Furnishing, and Services

3.1.1 Requirements

3.1.2 Services

3.1.3 Ventilation

3.2 Design and Layout

3.2.1 Sterile Handling Area

3.2.2 Laminar Flow Hoods

3.2.3 Service Bench

3.2.4 Quarantine and Containment

3.2.5 Incubation

3.2.6 Preparation Area

3.2.7 Storage

3.3 Disaster Management

4. Equipment and Materials.

4.1 Requirements of a Tissue Culture Laboratory

4.2 Sterile Working Area

4.2.1 Laminar-Flow BSC

4.2.2 Service Carts

4.2.3 Sterile Liquid Handling-Pipetting and Dispensing

4.2.4 Inverted Microscope

4.2.5 Camera and Monitor

4.2.6 Dissecting Microscope

4.2.7 Centrifuge

4.2.8 Cell Counting

4.3 Incubation and Culture

4.3.1 Incubator

4.3.2 Humid CO2 Incubator

4.3.3 Temperature Recorders

4.3.4 Roller Racks

4.3.5 Magnetic Stirrer

4.3.6 Culture Vessels

4.4 Preparation and Sterilization

4.4.1 Washup

4.4.2 Preparation of Media and Reagents

4.4.3 Sterilization

4.5 Storage

4.5.1 Consumables

4.5.2 Refrigerators and Freezers

4.5.3 Cryostorage Containers

4.5.4 Controlled-Rate Freezer

4.6 Supplementary Laboratory Equipment

4.6.1 Computers and Networks

4.6.2 Upright Microscope

4.6.3 Low-Temperature Freezer

4.6.4 Confocal Microscope

4.6.5 PCR Thermal Cycler

4.7 Specialized Equipment

4.7.1 Microinjection Facilities

4.7.2 Colony Counter

4.7.3 Centrifugal Elutriator

4.7.4 Flow Cytometer

Suppliers

5. Aseptic Technique

5.1 Objectives of Aseptic Technique

5.1.1 Risk of Contamination

5.1.2 Maintaining Sterility

5.2 Elements of Aseptic Environment

5.2.1 Laminar Flow

5.2.2 Quiet Area

5.2.3 Work Surface

5.2.4 Personal Hygiene

5.2.5 Reagents and Media

5.2.6 Cultures

5.3 Sterile Handling

5.3.1 Swabbing

5.3.2 Capping

5.3.3 Flaming

5.3.4 Handling Bottles and Flasks

5.3.5 Pipetting

5.3.6 Large-Volume Dispensing

5.3.7 Pouring

5.4 Standard Procedure

Protocol 5.1. Aseptic Technique In BSC

Protocol 5.2. Working On The Open Bench

5.4.1 Culture Flasks and Bottles

5.4.2 Petri Dishes and Multiwell Plates

Protocol 5.3. Handling Dishes or Plates

5.5 Apparatus and Equipment.

5.5.1 Refrigerators and Coldrooms

5.5.2 Incubators

Protocol 5.4. Cleaning Incubators

5.5.3 Boxed Cultures

5.5.4 Gassing with CO2

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

6.5.1 Operator

6.5.2 Equipment

6.5.3 Glassware and Sharp Items

6.5.4 Chemical Toxicity

6.5.5 Gases

6.5.6 Liquid Nitrogen

6.5.7 Burns

6.6 Fire

6.7 Ionizing Radiation

6.7.1 Ingestion

6.7.2 Disposal of Radioactive Waste

6.7.3 Irradiation from Labeled Reagents

6.7.4 Irradiation from High-Energy Sources

6.8 Biohazards

6.8.1 Levels of Biological Containment

6.8.2 Biological Safety Cabinets (BSCs)

6.8.3 Human Biopsy Material

6.8.4 Cell Lines

6.8.6 Genetic Manipulation

6.8.6 Disposal of Biohazardous Waste

6.8.7 Decontamination and Fumigation

6.9 Bioethics

6.9.1 Animal Tissue

6.9.2 Human Tissue

6.10 Quality Assurance

6.10.1 Procedures

6.10.2 Quality Control (QC)

6.10.3 Validation

6.10.4 Authentication

6.10.5 Provenance

6.10.6 Contamination

7. Culture Vessels and Substrates

7.1 The Substrate

7.1.1 Attachment and Growth

7.1.2 Common Substrate Materials

7.1.3 Alternative Substrates

7.2 Treated Surfaces

7.2.1 Substrate Coating

Protocol 7.1. Preparation of ECM

7.2.2 Feeder Layers

7.2.3 Nonadhesive Substrates

7.3 Choice of Culture Vessel

7.3.1 Cell Yield

7.3.2 Multiwell Plates

7.3.3 Flasks and Petri Dishes

7.3.4 High Yields

7.3.5 Suspension Culture

7.3.6 Venting

7.3.6 Sampling and Analysis

7.3.7 Uneven Growth

7.3.8 Cost

7.4 Specialized Systems

7.4.1 Permeable Supports

7.4.2 Three-Dimensional Matrices

Suppliers.

8. Defined Media and Supplements

8.1 D evelopment of Media

8.2 Physicochemical Properties

8.2.1 pH

Protocol 8.1. Preparation of pH Standards

8.2.2 CO2 and Bicarbonate

8.2.3 Buffering

8.2.4 Oxygen

Minireview 8.1. Hypoxic Cell Culture

8.2.5 Osmolality

8.2.6 Temperature

8.2.7 Viscosity

8.2.8 Surface Tension and Foaming

8.3 Balanced Salt Solutions

8.4 Complete Media

8.4.1 Amino Acids

8.4.2 Vitamins

8.4.3 Salts

8.4.4 Glucose

8.4.5 Organic Supplements

8.4.6 Hormones and Growth Factors

8.4.7 Antibiotics

8.5 Serum

8.5.1 Protein

8.5.2 Growth Factors

8.5.3 Hormones

8.5.4 Nutrients and Metabolites

8.5.5 Lipids

8.5.6 Minerals

8.5.7 Inhibitors

8.6 Selection of Medium and Serum

8.6.1 Serum Batch Reservation

8.6.2 Testing Serum

8.6.3 Heat Inactivation

8.7 Other Supplements

8.7.1 Amino Acid Hydrolysates

8.7.2 Embryo Extract

8.7.3 Conditioned Medium

8.8 Storage

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

9.4.1 Commercially Available Serum-Free Media

9.4.2 Serum Replacements

9.4.3 Serum-Free Subculture

9.4.4 Hormones

9.4.5 Growth Factors

9.4.6 Nutrients in Serum

9.4.7 Proteins and Polyamines

9.4.8 Viscosity

9.5 D evelopment of Serum-Free Medium

9.6 Selection of Serum-Free Medium

9.6.1 Cell or Product Specificity

9.6.2 Adaptation of Cell Lines to Serum-Free Media

9.7 Preparation of Serum-Free Medium

9.8 Animal Protein-Free Media

9.9 Conclusions

10. Preparation and Sterilization

10.1 Preparation of Reagents and Materials

10.2 Sterilization of Apparatus and Liquids

10.2.1 Dry-Heat Sterilization

10.2.2 Pressurized-Steam Sterilization.

10.2.3 Irradiation

10.2.4 Chemical Sterilization

10.2.5 Sterility Indicators

10.2.6 Filter Sterilization

10.3 Apparatus

10.3.1 Glassware

Protocol 10.1. Preparation and Sterilization of Glassware

10.3.2 Glass Pipettes

Protocol 10.2. Preparation and Sterilization of Glass Pipettes

10.3.3 Screw Caps

Protocol 10.3. Preparation and Sterilization of Screw Caps

10.3.4 Selection of Detergent

10.3.5 Miscellaneous Equipment

10.3.6 Reusable Sterilizing Filters

Protocol 10.4. Sterilizing Filter Assemblies

10.4 Reagents and Media

10.4.1 Water

Protocol 10.5. Preparation and Sterilization of Ultrapure Water (UPW)

10.4.2 Maintenance of Water Purifier

10.4.3 Balanced Salt Solutions

Protocol 10.6. Preparation and Sterilization of D-PBSA

10.4.4 Preparation and Sterilization of Media

Protocol 10.7. Preparation of Medium from 1× Stock

Protocol 10.8. Preparation of Medium from 10× Concentrate

Protocol 10.9. Preparation of Medium from Powder

10.5 Sterilization of Media

10.5.1 Autoclavable Media

10.5.2 Sterile Filtration

Protocol 10.10. Sterile Filtration with Syringe-Tip Filter

Protocol 10.11. Sterile Filtration with Vacuum Filter Flask

Protocol 10.12. Sterile Filtration with Small Inline Filter

Protocol 10.13. Sterile Filtration with Large Inline Filter

10.5.3 Serum

10.5.4 Preparation and Sterilization of Other Reagents

10.6 Control, Testing, and Storage of Media

10.6.1 Quality Control

10.6.2 Sterility Testing

10.6.3 Culture Testing of Medium and Serum

Protocol 10.14. Testing Medium by Plating Efficiency

Protocol 10.15. Testing Medium by Growth

10.6.4 Storage

11. Primary Culture

11.1 Initiation of a Primary Cell Culture

11.1.1 Proteases Used in Disaggregation

11.1.2 Common Features of Disaggregation.

11.2 Isolation of the Tissue.

Notes:

Previous ed.: 2010

Includes bibliographical references and index

Includes bibliographical references at the end of each chapters and index.

Description based on online resource; title from PDF title page (ebrary, viewed February 17, 2016).

ISBN:

1-118-87364-5

1-118-87337-8

OCLC:

935254494