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Cell culture techniques allow a variety of molecular and cell biological questions to be addressed, offering physiological conditions whilst avoiding the use of laboratory animals. In addition to basic techniques, a wide range of specialised practical protocols covering the following areas are included: cell proliferation and death, in-vitro models for cell differentiation, in-vitro models for toxicology and pharmacology, industrial application of animal cell culture, genetic manipulation and analysis of human and animal cells in culture.
List of contents
I General Methods.- 1 Cloning Animal Cells.- 2 Cell Culture and Diagnostic Virology.- 3 Screening for Mycoplasma Contamination in Animal Cell Cultures.- 4 Serum-Free Media.- 5 Screening Culture Media and Sera for Endotoxin Testing.- 6 Generation of Monoclonal Antibodies and Characterization of Novel Antibodies by Western Blotting and Immunocytochemistry.- 7 The Generation of Long-Term Human Antibody-Producing Cell Lines Using the Epstein-Barr Virus.- 8 Primary Culture.- II Cell proliferation and Death.- 9 Baculoviral Expression and Partial Purification of Cyclin/CDK Complexes.- 10 Methods for Studying Phosphoinositide Metabolism in Cultured Cells.- 11 Analysis of Cell Cycle and Cell Death Mechanisms.- III In Vitro Models for Cell Differentiation.- 12 Assessing Release of Secretory Products from Individual Cells.- 13 The Granulocyte/Macrophage Colony-Forming Unit Assay.- 14 Investigations into Cell Differentiation Using Cells in Culture.- 15 Cell Culture Models for Mononuclear Phagocytes.- 16 The Human Leukemia Cell Line HL-60 as a Cell Culture Model To Study Neutrophil Functions and Inflammatory Cell Responses.- 17 Cell Culture Models for Polarized Epithelial Monolayers.- 18 Ion Channels and Cell Signaling in Cell Cultures.- 19 Isolation, Cultivation and Differentiation of Lung Type II Epithelial Cells.- 20 Culture Systems for Hepatocytes for Use in Toxicology and Differentiation Studies.- IV In Vitro Models for Toxicology and Pharmacology.- 21 Cellular Models for In Vitro Toxicity Testing.- 22 Miniaturized In Vitro Methods in Toxicity Testing.- 23 Investigation of Anti-tumor Drug Accumulation and Efflux in Cell Culture.- 24 Current Status of Genetically Engineered In Vitro Systems for Biotransformation.- V Industrial Application of Animal Cell Culture.- 25 Scale-Up of Animal Cell Cultures.- 26 Purification of Monoclonal Antibodies Produced by Animal Cells in Culture.- 27 Biosafety Assessment of Cultured Animal Cells Used for Biopharmaceutical Production.- 28 Cytokine Cell Culture Assays.- VI Genetic Manipulation and Analysis of Human and Animal Cells in Culture.- 29 Antisense Methods in Cell Culture.- 30 Transfection with Ribozymes to Investigate the Role of Specific Gene Expression.- 31 Methods for Studying the Transcription and DNase I Hypersensitive Sites of Genes in Cells in Culture.- 32 Analysis of Specific and Differentially Expressed mRNAs in Cell Culture.- 33 Human Gene Therapy: Review and Outlook.
