Techniques in Somatic Cell Genetics

Somatic cell genetics is an exciting and rapidly expanding field of research. Since descriptions of the major experimental techniques in the field are scattered throughout various journals and other publications, there is a real need for a single reference source for both established investigators and students in the field. In addition, technical reports are frequently abridged such that many researchers are discouraged from attempting to adopt the appropriate methodology. This book, therefore, describes in detail the many recent technical advances in such areas of somatic cell genetics as transfer mediated by liposomes, erythrocyte ghosts, chromosomes, micro cells, mito­ chondria, and isolated nuclear DNA. These techniques have increased our understanding of the organization and regulation of eukaryotic cells. The production of antibiotic-resistant cell lines and their use in studying cytoplasmic inheritance are also included. Evidence for the cytoplasmic regulation of nuclear gene expression in eukaryotic cells is rapidly accumu­ lating following the characterization of cytoplasmic mutations. The produc­ tion of nuclear-coded mutations, their use in standard cell hybridization, and recent advances in techniques for fusing whole cells or cell components are also described.

Table of Contents:
1 Selection of Purine and Pyrimidine Nucleoside Analog Resistance in Mammalian Cells.- 2 Techniques for Using HAT Selection in Somatic Cell Genetics.- 3 Techniques for Decreasing the Toxicity of Polyethylene Glycol.- 4 The Use of Dimethyl Sulfoxide in Mammalian Cell Fusion.- 5 The Selection of Heterokaryons and Cell Hybrids Using the Biochemical Inhibitors Iodoacetamide and Diethylpyrocarbonate.- 6 Techniques for Enucleation of Mammalian Cells.- 7 Nonselective Isolation of Fibroblast Heterokaryons, Hybrids, and Cybrids by Flow Sorting.- 8 Techniques for Monitoring Cell Fusion: The Synthesis and Use of Fluorescent Vital Probes (R18, F18).- 9 Inheritance of Oligomycin Resistance in Tissue Culture Cells.- 10 Cytoplasmic Inheritance of Rutamycin Resistance in Mammalian Cells.- 11 Erythromycin Resistance in Human Cells.- 12 Cytoplasmic Inheritance of Chloramphenicol Resistance in Mammaliam Cells.- 13 The Influence of Cytoplast-to-Cell Ratio on Cybrid Formation.- 14 Transformation of Mitochondrially Coded Genes into Mammalian Cells Using Intact Mitochondria and Mitochondrial DNA.- 15 Mitochondrial Influences in Hybrid Cells.- 16 Shedding of Tumor Cell Surface Membranes.- 17 Production of Microcytospheres.- 18 Isolation and Characterization of Mitoplasts.- 19 Nuclear Transplantation with Mammalian Cells.- 20 Techniques for Purifying L-Cell Karyoplasts with Minimal Amounts of Cytoplasm.- 21 Techniques for Isolating Nuclear Hybrids.- 22 Monolayer Enucleation of Colcemid-Treated Human Cells and Polyethylene Glycol 400-Mediated Fusion of Microkaryoplasts (Microcells) with Whole Cells.- 23 Microcell-Mediated Chromosome Transfer.- 24 Techniques for Isolating Chromosome-Containing Minisegregant Cells.- 25 Techniques to Isolate Specific Human Metaphase Chromosomes.- 26 Techniques of Chromosome-Mediated Gene Transfer.- 27 Transfer of Macromolecules Using Erythrocyte Ghosts.- 28 Liposome-Mediated DNA Transfer.- 29 Techniques of DNA-Mediated Gene Transfer for Eukaryotic Cells.- 30 Viral-DNA Vectors in the Analysis of Mammalian Differentiation.- 31 Detection of Specific DNA Sequences in Somatic Cell Hybrids and DNA Transfectants.- 32 Microinjection Turns a Tissue Culture Cell into a Test Tube.- 33 Techniques of Somatic Cell Hybridization by Fusion of Protoplasts.- 34 Techniques for Chromosome Analysis.- 35 Genetic Analysis of Hybrid Cells Using Isozyme Markers as Monitors of Chromosome Segregation.- 36 Future Perspectives in Somatic Cell Genetics.