Bernice M. Martin

Tissue Culture Techniques

An Introduction

• Broschiertes Buch

Produktbeschreibung

ACKNOWLEDGMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Xl II INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I STERILITY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Aseptic Technique . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Physical manipulations - Use of the sterile cabinet (hood) Sterilization Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Heat - Radiation - Toxic gas - Filtration - Antibiotics Quality Control of Sterilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23 Routine labeling Suggested Readings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26 vi CONTENTS ROUTINE CELL CULTURE . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 Feeding Schedules and Media Components . . . . . . . . . . . . . . . . . . . . 29 General properties of media and salt solutions - water as a reagent· Establishingfeeding schedules Subcultivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Solutions and methods for adherent cells - Common enzyme solutions - Inoculating (seeding) the cultures Cell Enumeration and Cell Viability . . . . . . . . . . . . . . . . . . . . . . . . . . 54 Hemocytometer - Particle counter - Cell viability Putting Routine Methods to Work . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 Normal cell growth characteristics Detecting and Disposing of Contamination . . . . . . . . . . . . . . . . . . . . 66 Bacteria and fUngi - Fungi -Mycoplasma - Viruses - Dealing with contamination Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73 Inadequate cell growth - Recurrent contamination - When to call your vendor Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80 Biological hazards - Chemical hazards Suggested Readings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Problem Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 EXPERIMENTS IN CULTURE . . . . . . . . . . . . . . . . . . . . . . . . . . 91 II Alterations of the Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 Serum - Treatments of serum - Plasma-derived serum - Serum-free and low-protein media Substrata. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101 Coatingplasticware with solutions - Alterations with polymers - Using cells to coat the plasticware - Culturing cells on microcarriers Altering the Environment. . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106 Temperature changes - Gaseous changes Problem Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110 CONTENTS vii PRIMARY CELL CULTURE. . . . . . . . . . . . . . . . . . . . . . . . . . . . 113 Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114 Dissection - Enzymatic dissociation methods- Nonenzymatic isolation - Purification of cell suspensions - Consideringyield and survival Chatacterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Produktdetails

• Verlag: Birkhäuser Basel / Springer, Basel
• 1994.
• Seitenzahl: 264
• Erscheinungstermin: 1. August 1994
• Englisch
• Abmessung: 254mm x 178mm x 15mm
• Gewicht: 540g
• ISBN-13: 9780817636432
• ISBN-10: 0817636439
• Artikelnr.: 21586122

Inhaltsangabe

1 Introduction.- 2 Sterility.- Aseptic Technique.- Sterilization Methods.- Quality Control of Sterilization.- Suggested Readings.- Exercises.- 3 Routine Cell Culture.- Feeding Schedules and Media Components.- Subcultivation.- Cell Enumeration and Cell Viability.- Putting Routine Methods to Work.- Detecting and Disposing of Contamination.- Troubleshooting.- Safety.- Suggested Readings.- Problem Set.- Exercises.- 4 Experiments in Culture.- Alterations of the Media.- Substrata.- Altering the Environment.- Problem Set.- Exercises.- 5 Primary Cell Culture.- Isolation.- Characterization.- Exercise.- 6 Cell Preservation.- Freezing the Cells.- Thawing the Cells.- Problem Set.- 7 Cell Cloning.- Isolation.- Assisting Clonal Survival.- Problem Set.- 8 Culture Changes.- The Loss of Differentiated Phenotype.- Immortality and Aging.- Cell Transformation.- Transfection (Gene Transfer).- Suggested Readings.- Problem Set.- Exercises.- 9 Information for New Cell Studies.- Searching in the Literature.- Using Advertising and Catalogs.- References, Bibliographies, and Databases.- Appendices.- A The Cell Cycle.- Suggested Readings.- B Media and Salt Solutions.- Media.- Salt Solutions.- C Vendors.- D Glossary.- E Answers to Problem Sets and Exercises.