Animal Cell Culture, 1990 Methods in Molecular Biology Series, Vol. 5

For many years I performed tissue culture in large scientific insti- tions that had a great deal of infrastructure. When I set up a tissue l- oratory outside such an infrastructure, however, I found there was a shortage of easily accessible information about the basic needs, reagents, and techniques for establishing such a facility. Much had to be done by trial and error or gleaned from originalpapers. Consequently, I felt that a methods book covering a wide variety of techniques from basic culture to the most sophisticated cell analysis would be a very valuable addition to the scientific literature. In the interim, several useful books (listed in Chapter I of this volume) did appear, but none entirely fitted the bill and some are now somewhat dated. Then, in 1984, the first of the Methods in Molecular Biology volumes from Humana Press was published with its step-by-step recipe approach. This format appealed to me, and so I c- tacted John Walker, the series editor, about including cell culture in this series. The result was that we embarked upon a single volume covering both plant and animal cell culture. Such was the richness of the material that this project soon divided itself into separate volumes on animal cell (Volume 5) and plant cell (Volume 6) culture. In this volume (Volume 5), therefore, we have aimed to describe a variety of basic techniques and culture conditions for a range of cell types.

Basic Cell Culture.- Establishment, Maintenance, and Cloning of Human Primary Cell Strains.- Aging of Cultured Human Skin Fibroblasts.- Separation and Maintenance of Primary T and B Lymphocytes.- Establishment of Lymphoblastoid Cell Lines.- Scale-Up of Suspension and Anchorage-Dependent Animal Cells.- Mycoplasma Detection.- Short-Term Chorionic Villi and Amniotic Fluid Cultures.- Keratinocyte Culture.- Establishment of Mouse Epithelial Lung Cell Strains and Cell Lines.- Culture of Human Brain Tumors on an Extracellular Matrix Derived from Bovine Corneal Endothelial Cells and Cultured Human Glioma Cells.- Embryonic Rodent Brain Cells in Culture.- Human Thyroid Epithelial Cells.- Preparation of Isolated Rat Liver Hepatocytes.- Primary Cultures of Rat Hepatocytes.- Preparation of Isolated Periportal or Perivenous Hepatocytes from Rat Liver.- Primary Kidney Cells.- Adipocytes.- Tissue Culture of Skeletal Muscle.- Derivation and Maintenance of Embryonic Stem Cell Cultures.- Defining Hormone and Matrix Requirements for Differentiated Epithelia.- Culturing Primitive Hemopoietic Cells.- High Proliferative Potential Colony Forming Cells.- Murine Bone Marrow-Derived Macrophages.- Long-Term B Lymphoid Cultures from Murine Bone Marrow.- CFU-C in Agar.- Human Long-Term Bone Marrow Culture.- In Vitro Clonal Culture of Human Hematopoietic Progenitor Cells.- Flow Sorting for Isolating CFU-E.- Production of Human and Murine Eosinophils In Vitro and Assay for Eosinophil Differentiation Factors.- Clonogenic Assays for Hematopoietic and Tumor Cells Using Agar-Containing Capillaries.- Cytogenetic Techniques for Human Leukemias.- Time-Lapse Cinemicroscopy.- Transmission and Scanning Electron Microscope Preparation of Whole Cultured Cells.- Double Indirect-Immunofluorescent Labeling of Cultured Cells.- Gene Mapping to Chromosomes by Hybridization in Situ.- In Situ Hybridization with Radiolabeled cRNA Probes, Using Tissue Sections and Smears.- Immunogold Labeling for Electron Microscopy.- Human Chromosome Analysis and Sorting.- Flow Cytometry.- Fluorescence-Activated Cell Sorting According to Receptor Density.- Transfection and Transformation of Human Thyroid Epithelial Cells.- Expression of Foreign Genes in Cultured Insect Cells Using a Recombinant Baculovirus Vector.- Chloramphenicol Acetyltransferase as a Reporter in Mammalian Gene Transfer.- Immunizing Schedules for Hybridoma Production.- Fusion Protocol for the Production of Mouse Hybridomas.- Cloning of Hybridomas.- Enzyme-Linked Immunosorbent Assay for Screening of Antibodies in Hybridoma Supernatants.- Cryopreservation of Hybridomas.- Isotype Determination of Monoclonal Antibodies by Immunodiffusion.- Isoelectric Focusing and Immunofixation of Monoclonal Antibodies.- Large-Scale Production and Storage of Monoclonal Antibodies and Hybridomas.- Purification of Murine Monoclonal Antibodies.- Rat x Rat Hybridomas.- Monoclonal Antibodies Against Glycosphingolipids (GSLs)-Gangliosides.