Beschreibung
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.