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AuthorMartin, Bernice M. author
TitleTissue Culture Techniques [electronic resource] : An Introduction / by Bernice M. Martin
ImprintBoston, MA : Birkhรคuser Boston : Imprint: Birkhรคuser, 1994
Connect tohttp://dx.doi.org/10.1007/978-1-4612-0247-9
Descript 260 p. online resource

SUMMARY

ACKNOWLEDGMENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Xl II INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I STERILITY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Aseptic Technique . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 Physical manipulations โ{128}ข Use of the sterile cabinet (hood) Sterilization Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Heat โ{128}ข Radiation โ{128}ข Toxic gas โ{128}ข Filtration โ{128}ข 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 โ{128}ข water as a reagentยท Establishingfeeding schedules Subcultivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 Solutions and methods for adherent cells โ{128}ข Common enzyme solutions โ{128}ข Inoculating (seeding) the cultures Cell Enumeration and Cell Viability . . . . . . . . . . . . . . . . . . . . . . . . . . 54 Hemocytometer โ{128}ข Particle counter โ{128}ข Cell viability Putting Routine Methods to Work . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 Normal cell growth characteristics Detecting and Disposing of Contamination . . . . . . . . . . . . . . . . . . . . 66 Bacteria and fUngi โ{128}ข Fungi โ{128}ข Mycoplasma โ{128}ข Viruses โ{128}ข Dealing with contamination Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73 Inadequate cell growth โ{128}ข Recurrent contamination โ{128}ข When to call your vendor Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80 Biological hazards โ{128}ข Chemical hazards Suggested Readings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Problem Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 EXPERIMENTS IN CULTURE . . . . . . . . . . . . . . . . . . . . . . . . . . 91 II Alterations of the Media . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 Serum โ{128}ข Treatments of serum โ{128}ข Plasma-derived serum โ{128}ข Serum-free and low-protein media Substrata. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101 Coatingplasticware with solutions โ{128}ข Alterations with polymers โ{128}ข Using cells to coat the plasticware โ{128}ข Culturing cells on microcarriers Altering the Environment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 106 Temperature changes โ{128}ข Gaseous changes Problem Set . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110 Exercises . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110 CONTENTS vii PRIMARY CELL CULTURE. . . . . . . . . . . . . . . . . . . . . . . . . . . . 113 Isolation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114 Dissection โ{128}ข Enzymatic dissociation methods โ{128}ข Nonenzymatic isolation โ{128}ข Purification of cell suspensions โ{128}ข Consideringyield and survival Chatacterization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .


CONTENT

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


Medicine General practice (Medicine) Medicine & Public Health General Practice / Family Medicine



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