Cell Culture for Biochemists

Acknowledgements. Chapter 1. Introduction. 1.1 Background. 1.2 Some advantages. 1.3 Applications.1.4 Animal cell biotechnology. Chapter 2. Characteristics of cultured cells. 2.1 Types of cells. 2.2 Primary cells and transformation. 2.3 Growth control. 2.4 Attachment and Spreading. 2.5 Growth factors. 2.6 Differentiated functions in cell cultures. Chapter 3. Culture vessels. 3.1 Design of culture vessels. 3.2 Small scale cultures. 3.3 Intermediate scale cultures. 3.4 Large scale cultures. 3.5 Suspension cultures. 3.6 Microcarriers. 3.7 Air lift systems. 3.8 Bioreactors. Chapter 4. Subculturing. 4.1 Dissociation techniques. 4.2 Subculture of a cell monolayer. 4.3 Subculture of cells growing in suspension. 4.4 Protocol for setting up microcarrier cultures 4.5 Subculture of cells growing on microcarriers. 4.6 The growth cycle. Chapter 5. Cell culture media. 5.1 Introduction. 5.2 Balanced salt solutions. 5.3 Eagle's medium. 5.4 More complicated media. 5.5 Simple media with unspecified additives. 5.6 Antibiotics. 5.7 Serum. 5.8 Serum-free media. 5.9 Media for culture of insect cells. 5.10 Media for culture of plant cells. Chapter 6. Primary cells. 6.1 Introduction. 6.2 Lymphocytes. 6.3 Human skin biopsies. 6.4 Mouse or rat embryo cultures. 6.5 Chick embryo cells. 6.6 Chick embryo liver cells. 6.7 Rat hepatocytes. 6.8 Primary kidney cells. 6.9 Endothelial cells. 6.10 Mammary epithelial cell cultures. 6.11 Colonic epithelial cells. 6.12 Rat or chick skeletal muscle cells. 6.13 Mouse macrophage cultures. 6.14 Ascites cells. 6.15 Dipteran cell culture. Chapter 7. Techniques. 7.1 Cell cloning and plating efficiency. 7.2 Cell counting procedures. 7.3 Storage of cells. 7.4 Karyotyping. 7.5 Cell transfection. 7.6 Cell visualisation. 7.7 Sub-cellular fractionation. Chapter 8. Glassware preparation and sterilisation techniques. 8.1 General. 8.2 Sterilisation by heat. 8.3 Sterilisation by filtration. Chapter 9. Contamination. 9.1. Bacterial contamination. 9.2 Sterility checks. 9.3 Analysis of bacterial contamination. 9.4 Airborne contamination. 9.5 Antibiotics. 9.6 Disposal of contaminated material. 9.7 Mycoplasmas. 9.8 Testing for mycoplasma 9.9 Elimination of mycoplasmas. 9.10 Viral contamination. Chapter 10. The cell cycle. 10.1 Description. 10.2 Mitosis. 10.3 S-phase. 10.4 Control of the cell cycle. 10.5 Distribution of cells around the cycle. 10.6 Growth fraction. 10.7 Cell cycle analysis. Chapter 11. Cell synchronisation. 11.1 Intoduction. 11.2 Selection of mitotic cells. 11.3 Selective killing of cells in particular phases. 11.4 Selection of cells by size. 11.5 Synchronisation by subculture. 11.6 Serum deprivation. 11.7 Isoleucine starvation. 11.8 Blockade of S-phase. 11.9 Procedure for inducing synchrony at the G1/S interphase. 11.10 Synchronisation in G2. 11.11 Synchronisation in M. Chapter 12. Use of radioactive isotopes in cell culture. 12.1 Estimation of rates of DNA synthesis. 12.2 Estimation of rates of RNA and protein synthesis. 12.3 Autoradiography. 12.4 Labelling with bromodeoxyuridine. 12.5 DNA repair. Chapter 13. Cell mutants and cell hybrids. 13.1 Auxotrophic mutants. 13.2 Selection of mutants. 13.3 Temperature sensitive mutants. 13.4 Replica plating of animal cells. 13.5 Somatic cell hybridisation. 13.6 Myeloma culture and monoclonal antibody production. 13.7 Methods of fusion. 13.8 Cell communication. Chapter 14. Viruses. 14.1 Introduction. 14.2 Virus production. 14.3 Virus detection. 14.4 Production and testing of viral vaccines. 14.5 Viral transformation of cells. Chapter 15. Differentiation in cell cultures. 15.1 Erythroid differentiation of Friend cells. 15.2 Skin and keratinocytes. 15.3 Teratocarcinoma cells. 15.4 Differentiation of muscle cells. 15.5 Differentiation of adipose cells. 15.6 Differentiated hepatocytes. Chapter 16. Appendices. References. Subject Index.