Tissue Culture

Tissue Culture: Methods and Applications presents an overview of the procedures for working with cells in culture and for using them in a wide variety of scientific disciplines. The book discusses primary tissue dissociation; the preparation of primary cultures; cell harvesting; and replicate culture methods. The text also describes protocols on single cell isolations and cloning; perfusion and mass culture techniques; cell propagation on miscellaneous culture supports; and the evaluation of culture dynamics. The recent techniques facilitating microscopic observation of cells; cell hybridization; and virus propagation and assay are also encompassed. The book further tackles the production of hormones and intercellular substances; the diagnosis and understanding of disease; as well as quality control measures. Scientists and professionals interested in methodology per se will find the book invaluable.

List of Contributors

Preface

Section I. Primary Tissue Dissociation

Chapter 1. Pronase

Chapter 2. Trypsin

A. Mammalian Tissues

B. High Yield Method for Kidney Tissue

C. Marine Teleost Fish Tissues

D. Amphibian Tissues

Chapter 3. Collagenase Treatment of Chick Heart and Thyroid

Chapter 4. Sequential Enzyme Treatment of Mouse Mammary Gland

Chapter 5. Nonenzymatic Dissociations

A. Leukocyte Cell Separation on Glass

B. Specific Cell Fractionation on Chemically Derivatized Surfaces

Section II. Preparation of Primary Cultures

Chapter 1. Human Skin Fibroblasts

Chapter 2. Fetal Human Diploid Cells

Chapter 3. Human Mammary Tumors

Chapter 4. Human Glia Cells

Chapter 5. Human Liver Cells

Chapter 6. Human Blood Leukocytes

Chapter 7. Amniocentesis

Chapter 8. Human Lymph Nodes

Chapter 9. Human Biopsy Material from Genetic Abnormalities

Chapter 10. Mammalian Myocardial Cells

Chapter 11. Mammalian Glomerular Cells

Chapter 12. Organotypic Mammalian Central and Peripheral Nerve Tissue

Chapter 13. Marmoset Bone Marrow

Chapter 14. Monkey Kidney Cells

Chapter 15. Rabbit Kidney and Skin

Chapter 16. Rat Skeletal Muscle Cells

Chapter 17. Rat Pituitary Explants

Chapter 18. Chick Embryo Cells

Chapter 19. Frog Embryos ( Haploid Lines)

Chapter 20. Adult Amphibian Tissues and Leukocytes

Chapter 21. Marine Teleost Fish Tissues

Chapter 22. Fish Lymphocytes and Blastogenesis

Chapter 23. Freshwater Fishes

Chapter 24. Goldfish Tissues

Chapter 25. Reptiles

Chapter 26. Dipteran Embryos and Larvae (Diploid Lines)

Chapter 27. Cockroach and Grasshopper Embryo Tissue

Chapter 28. Plants: Haploid Tissue Cultures

Chapter 29. Aspen Callus

Chapter 30. Plant Embryos

Chapter 31. Somatic Plant Cells

Chapter 32. Roots

Chapter 33. Leaves

Section III. Cell Harvesting

Chapter 1. Pronase

Chapter 2. Trypsinization of Diploid Human Fibroblasts

Chapter 3. Transplantable Rat Tumors and Cold Temperature Release

Chapter 4. Cells at Defined Stages of the Cell Cycle

A. Selection of Mitotic Cells from Monolayer Culture

B. Sedimentation in Sucrose and Ficoll Gradients of Cells Grown in Suspension Culture

Chapter 5. Processing Cells for Enzyme Assays

Section IV. Replicate Culture Methods

Chapter 1. Monolayer Cultures

Chapter 2. Plant Cell Suspension Cultures

Chapter 3. Subculturing Human Diploid Fibroblast Cultures

Chapter 4. Apparatus for Changing Tissue Culture Media

Chapter 5. Single "Subculture" in Situ Using Glass Beads

Chapter 6. Maintenance of Diploid Fibroblast Cultures as Nonmitotic Populations

Section V. Single Cell Isolations and Cloning

Chapter 1. Animal Cells

A. Capillary Techniques

B. Microdrop Techniques

Chapter 2. Plant Cells

Chapter 3. Dilution Plating and Nutritional Considerations

A. Animal Cells

B. Plant Cells

Chapter 4. Dilution Plating on Coverslip Fragments

Chapter 5. Microtest Plates

A. Single Cell Clones

B. Replica Plating

Chapter 6. Low Oxygen Tension Technique

Chapter 7. Soft Agar Techniques

Section VI. Perfusion and Mass Culture Techniques

Chapter 1. Dual-Rotary Circumfusion System

Chapter 2. Automated Single Rose Chamber for Cardiac and Other Cells

Chapter 3. Culture Dish Perfusion with Cinemicrography

Chapter 4. Sykes-Moore Chamber

Chapter 5. Culture of the Lens (33°)

Chapter 6. Cardiac Cells in a Capillary Tube

Chapter 7. Artificial Capillaries: An Approach to Tissue Growth in Vitro

Chapter 8. Replicate Roller Bottles

Chapter 9. Replicate Tube Chambers for Bones

Chapter 10. Multisurface Stacked Plate Propagators

Chapter 11. Bulk Culture of Cell Monolayers

Chapter 12. Spin Filter Device for Suspension Cultures

Chapter 13. Automation of Continuous Suspension Cultures by Means of a Nephelostat

Chapter 14. A Chemostat for Single Cell Cultures of Higher Plants

Section VII. Cell Propagation on Miscellaneous Culture Supports

Chapter 1. Collagen-Coated Cellulose Sponge

Chapter 2. Microcarrier Cultures of Animal Cells

Chapter 3. Polyester Sheeting

Chapter 4. Protein Polymers as a Substratum for the Modulation of Cell Proliferation in Vitro

Chapter 5. Titanium Disks

Chapter 6. Filter Paper Supports for Plant Cultures

Section VIII. Evaluation of Culture Dynamics

Chapter 1. Hemocytometer Counting

Chapter 2. Electronic Enumeration and Sizing of Cells

A. Primary Tissue Cells

B. Tissue Culture Cells

Chapter 3. Dye Exclusion Tests for Cell Viability

Chapter 4. Quantitative Methods for Measuring Cell Growth and Death

Chapter 5. Cell Cycle Analysis

A. Mammalian Cells

B. Control and Analysis of the Mitotic Cycle in Cultured Plant Root Meristems

Section IX. Recent Techniques Facilitating Microscopic Observation of Cells

Chapter 1. Contrast Methods in the Microscopy of Living Tissue

Chapter 2. Rapid Embedding of Cell Culture Monolayers and Suspensions

Chapter 3. Embedding in Situ

Chapter 4. Vertical Sectioning

A. Cells on Millipore Filters

B. Cells in Plastic Flasks

C. Cells Grown on Decalcified Eggshell Membrane with Special Reference to Electron Microscopic Study

Chapter 5. Hydraulically Operated Microscope Stage

Chapter 6. Culture Dish for High Resolution Microscopy

Section X. Cell Hybridization

Chapter 1. Somatic Hybridization in Studies of Heredity of Cell Malignancy

Chapter 2. Production and Characterization of Proliferating Somatic Cell Hybrids

Chapter 3. Plant Protoplast Culture

Section XI. Virus Propagation and Assay

Chapter 1. Microculture Procedures

A. Vesicular Stomatitis Virus

B. Simian Virus 40

C. Antibody Neutralization Surveys

Chapter 2. Quantal and Enumerative Titration of Virus in Cell Cultures

Chapter 3. Diagnosis of Virus-Infected Cultures with Fluorescein-Labeled Antisera

Chapter 4. Rapid Method for Detection of Budding Virus by Electron Microscopy

Chapter 5. Monitoring for Presence of Oncogenic Virus in Tissue Culture

Chapter 6. Tobacco Mosaic Virus in Plant Tissue Culture

Section XII. Production of Hormones and Intercellular Substances

Chapter 1. Growth Hormone and Prolactin from Rat Pituitary Tumor Cells

Chapter 2. Enhancement of Growth Hormone Production by Cortisol

Chapter 3. Human Chorionic Gonadotropin

Chapter 4. Parathyroid Hormone and Calcitonin

Chapter 5. Measurement of Collagen Synthesis

Chapter 6. Evaluation of Milk Protein Synthesis

Chapter 7. Interferon Yield versus Cell Genotype

Chapter 8. Immunoglobulins

Section XIII. Diagnosis and Understanding of Disease

Chapter 1. Some Cell Culture Procedures in Diagnostic Medical Virology

Chapter 2. Diagnostic Use of Cell Cultures Initiated from Amniocentesis

Chapter 3. Expressions of Citrullinemia in Skin Fibroblast Cultures

Chapter 4. The Hurler Cell in Culture

Chapter 5. Characteristics of Fibroblasts Cultured from Patients with Galactosemia or Galactokinase Deficiency

Chapter 6. Evaluation of Amino Acid Metabolism in Maple Syrup Urine Disease

Chapter 7. Malignant Transformation of Cells in Culture Using Oncogenic Chemicals

Chapter 8. Malignant Transformation of Cells in Culture

Chapter 9. Some in Vitro Models of Normal and Malignant Cell Interaction

Chapter 10. Growth and Isolation of Nutritional Variants

Section XIV. Quality Control Measures

Chapter 1. Glassware Preparation, Sterilizations, and Use of Laminar Flow Systems

Chapter 2. Purification and Standardization of Water for Tissue Culture

Chapter 3. A Surveillance Procedure Applied to Sera

Chapter 4. Preparation of Sera for Nutritional Studies

Chapter 5. Sample Preparations of Media

A. Vertebrate Cells

B. Invertebrate Cells

C. Plant Cultures

Chapter 6. Determination and Survey of Osmolality in Culture Media

Chapter 7. Control of Culture pH with Synthetic Buffers

Chapter 8. Freezing, Storage, and Recovery of Cell Stocks

Chapter 9. Detection, Elimination, and Prevention of Bacteria and Fungi in Tissue Cultures

Chapter 10. Screening Tissue Cultures for Mycoplasma Infections

Chapter 11. Mycoplasma Contamination of Cell Cultures: Incidence, Source, Prevention, and Problems of Elimination

Chapter 12. Elimination of Pathogens from Shoot Tip Cultures

Chapter 13. Contamination of Tissue Culture and Its Cure: Some Examples

Chapter 14. Serological Identifications of Cells in Culture

A. Animal Cells by Fluorescent-Labeled Antibody

B. Poikilotherm Cell Identity by Cytotoxic Antibody Test

C. Invertebrate Cell Cultures

D. Detection of HL-A Antigens on Cultured Normal Human Diploid Fibroblasts

Chapter 15. Karyology of Cells in Culture

A. Preparation and Analysis of Karyotypes and Idiograms

B. Harvesting Human Leukocyte Cultures for Chromosome Cytology

C. Constitutive Heterochromatin (C-Bands)

D. Fluorescent Banding of Chromosomes (Q-bands)

E. Trysin Technique to Reveal G-Bands

F. Characteristics of Insect Cells

G. Characteristics of Plant Cells

Chapter 16. Biochemical Identification of Cells in Culture

A. Human Cell Lines by Enzyme Polymorphism

B. Enzymatic "Fingerprinting"

Chapter 17. Introduction to Cytoenzymological Methods and Isozymes

Chapter 18. Transport and Shipping of Cultures, Including International Regulations

Author Index

Subject Index