Handbook of Experimental Pharmacology Continuation of Handbuch der experimentellen Pharmakologie

Vol. 57

Editorial Board

G. V. R. Born, London' A. Farah, Rensselaer, New York H. Herken, Berlin· A. D. Welch, Memphis, Tennessee

Advisory Board S. Ebashi· E. G. Erdos' V. Erspamer' U. S. von Euler' W. S. Feldberg G. B. Koelle· O. Krayer' M. Rocha e Silva· J. R. Vane· P. G. Waser

Tissue Growth Factors

Contributors

1. Abrahm . R Baserga . G. Carpenter· D. R. Clemmons M. 1. Cline . V. 1. Cristofalo . D. D. Cunningham P. Datta· Ch. M. Dollbaum . J. H. Fitchen . E Grinnell P. M Gu1lino . R G. Ham . L. Harel . 0. H. Iversen M. 1. Koroly . D. Metcalf· B. A Rosner· R Ross G. Rovera . H. S. Smith· J. J. Van "\\yk . M. Young

Editor

R. Baserga

Springer-Verlag Berlin Heidelberg New York 1981

Professor RENATO BASERGA, M.D. Senior Investigator, Fels Research Institute and Department of Pathology Temple University School of Medicine. Health Sciences Center Philadelphia, PA 19l40jUSA

With 47 Figures

ISBN -13 :978-3-642-67988-9 e-ISBN-13 :978-3-642-67986-5 DOl: 10.1007/978-3-642-67986-5

Library of Congress Cataloging in Publication Data. Main entry under title: Tissue growth factors. (Handbook of experimental pharmacology; v.57) Bibliography: p. Includes index. I. Tissues - Growth. 2. Growth promoting substances. 3. Cell proliferation. I. Abraham, J. (Janet) II. Baserga, Renato. Ill. Series. [DNLM: I. Cell division. 2. Growth substances. 3. Peptides. WI HA51L v. 57/QH 511 T616] QP905.H3 vol. 57 [QP88] 615'.ls [599.08'2] 80-14574AACR2 ISBN-I 3:978-3-642-67988-9

This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned specifically those of translation, reprinting, re-use of illustrations, broadcasting, reproducing by photocopying machine or similar means, and storage in data banks. Under § 54 of the German Copyright Law where copies are made for other than private use a fee is payable to "Verwcrtungsgesellschaft Wort", Munich.

(g by Springer-Verlag Berlin Heidelberg 1981 Softcover reprint of the hardcover I st edition

The use of registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use.

Typesetting, printing, and bookbinding: Briihlsche Universitiitsdruckerei, Giessen. 2122/3130-543210

List of Contributors

Dr. JANET ABRAHM, Hematology-Oncology Division, Department of Internal Medicine, University of Pennsylvania, Philadelphia, PA 19140(USA

Professor R. BASERGA, M.D., Senior Investigator, Fels Research Institute and Department of Pathology, Temple University School of Medicine, Health Sciences Center, Philadelphia, PA 19140jUSA

Professor G. CARPENTER, Departments of Biochemistry and Medicine, Vanderbilt University, School of Medicine, Nashville, TN 37232(USA

Professor D. R. CLEMMONS, M.D., University of North Carolina, Department of Medicine and Pediatrics, Clinical Sciences Building 229H, Chapel Hill, NC27514(USA

Professor M. J. CLINE, M.D., Department of Medicine, Center for the Health Sciences, University of California, Los Angeles, CA 90024(USA

Professor V. J. CRISTOFALO, Ph.D., The Wistar Institute, 36th Street at Spruce, Philadelphia, PA 19104(USA

Professor D. D. CUNNINGHAM, Department of Microbiology, College of Medicine, University of California, Irvine, CA 92717 (USA

Professor P. DATTA, Department of Biological Chemistry, University of Michigan, Medical School, P.O.B. 034, Ann Arbor, MI48109(USA

Professor CH. M. DOLLBAUM, Peralta Cancer Research Institute, 3023 Summit Street, Oakland, CA 94609(USA

J. H. FITCHEN, M.D., Assistant Professor of Medicine, Department of Medicine, UCLA School of Medicine, Center for the Health Sciences, Los Angeles, CA 90024jUSA

Professor F. GRINNELL, Department of Cell Biology, University of Texas, Health Sciences Center at Dallas, Southwestern Medical School, Dallas, TX 75235jUSA

Dr. P. M. GULLINO, Department of Health, Education, and Welfare, Public Health Service, National Institutes of Health, Bethesda, MD 200 14(USA

Professor R. G. HAM, University of Colorado, Department of Molecular, Cellular, and Developmental Biology, Biosciences Building, Boulder, CO 80309(USA

VI List of Contributors

Dr. LOUISE HAREL, Institut de Recherches, Scientifiques Sur Le Cancer, 7, Rue Guy-Mocquet, F-94800 Villejuif

Professor O. H. IVERSEN, Universitet I Oslo, Institutt for Patologi, Rikshospitalet, N-Oslo 1. McArdle Laboratory for Cancer Research, University of Wisconsin, Madison, WI53706jUSA

Professor MARY J. KOROLY, Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, Gainesville, FL 32610jUSA

D. METCALF, M.D., Head Cancer Research Unit, The Walter and Eliza Hall, Institute of Medical Research, Royal Melbourne Hospital, AUS-Victoria 3050

Dr. B. A. ROSNER, The Wistar Institute of Anatomy and Biology, 36th Street at Spruce, Philadelphia, PA 19104jUSA

Professor R. Ross, University of Washington, School of Medicine, Department of Pathology and Biochemistry, Seattle, WA 98195jUSA

Professor G. ROVERA, The Wi star Institute of Anatomy and Biology, 36th Street at Spruce, Philadelphia, PA 19104jUSA

HELENE S. SMITH, Ph.D., Assistant Director, Peralta Cancer Research Institute, 3023 Summit Street, Oakland, CA 94609jUSA

Professor J. J. VANWYK, Departments of Medicine and Pediatrics, University of North Carolina, School of Medicine, Clinical Sciences, Building 229H, Chapel Hill, NC 27514jUSA

Professor M. YOUNG, Department of Biochemistry and Molecular Biology, University of Florida, College of Medicine, Gainesville, FL 32610jUSA

Preface

From a logical point of view, cell division is regulated by the environment and by the ability of the cell to respond to the environmental signals. The terminology of the cell cycle, the elaborate mathematical models, and the kinetic analyses are all convenient notations and descriptions of the behavior of populations of cells. However, they tell us very little about the fundamental molecular mechanisms that control cell proliferation. Stated in other terms, what controls cell reproduction are growth factors in the environment and genes and gene products inside the cell or at its surface.

This book examines the aforementioned growth factors, the study of which has made very rapid progress in the past few years. The selection of topics has been influenced by logistic considerations, but the book, as a whole, gives a broad survey of the state of the art of this exciting field. For this, thanks are due to the contributors, who have given much time to the preparation of the manuscripts and have met the deadline with a punctuality that is uncommon among biomedical scientists.

I would also like to thank Ms. NORA PERRETT and the staff of Springer-Verlag for their help in editing the manuscripts and in preparing the production of the book.

Philadelphia R. BASERGA

Contents

CHAPTER 1

Introduction to Cell Growth: Growth in Size and DNA Replication R. BASERGA

A. Introduction. . . . . . . . . . . . 1 B. Doubling of Size in Growing Cells . . 1 C. Replication of DNA in Growing Cells 3 D. Independence of Signal to Grow in Size from Signal to Replicate DNA 6 E. Mechanism of Action of Growth Factors 7 References . . . . . . . . . . . . . . . . . . . . . . . . . . .. 8

CHAPTER 2

Survival and Growth Requirements of Nontransformed Cells. R. G. HAM

A. Definitions . . . . . . . . . . . . . . . . . . . . 13 I. Normal, Nontransformed, and Transformed Cells. . . . . 13

II. Survival and Survival Requirements. . . . . . . . . . . 14 III. Cell Cycle, Growth, Division, Multiplication, and Proliferation 14 IV. Nutrient, Growth Requirement, Growth Factor, Mitogen,

and Hormone. . . . . . . . . . . . . . . . . . . 15 B. Assay Systems for the Measurement of Growth Requirements . 16

I. Background . . . . . . . . . . . . . . . . . . . . 16 II. Inadequacy for Nontransformed Cells of Classic Assay Systems. 17

1. Evolutionary Adaptation of Permanent Lines . . . . . . . 17 2. Role of Serum in the Multiplication of Nontransformed Cells 18 3. Problems Awaiting Solution for Nontransformed Cells 19

III. Holistic Approach to Cellular Growth Requirements 20 IV. Development of Specific Assays for Individual Growth

Requirements . . . . . . . . . . . . . . . . . . 21 1. Background Media Lacking a Single Growth-Promoting

Substance . . . . . . . . 22 2. First Limiting Factor . . . 22 3. Replaceable Requirements. 22 4. Sequential Depletion . . . 23 5. Systematic Testing of Suspected Growth-Promoting Substances 23

x Contents

V. Types of Measurement and Analysis of Data 23 1. Short-Term Vs Long-Term Multiplication 24 2. Sparse Vs Dense Cultures 24 3. DNA Synthesis Vs Increase in Cell Number 25 4. Analysis of Data Based on Total Multiplication 26 5. Analysis Based on Rate of Multiplication 27 6. Multiplication Rate Kinetics . 28

VI. Systematic Analysis of Growth Requirements . 28 C. Requirements for Survival and Growth of Nontransformed Cells 33

I. Transformed Vs Nontransformed Cells 33 II. Requirements Related to Subculturing and Cellular Attachment. 35

1. Anchorage Dependence . 35 2. Neutralization of the Dispersing Agent 35 3. Low Temperature Trypsinization . 36 4. Attachment and Spreading Factors . 37 5. The Culture Surface 38 6. Artificial Substitutes for the Basement Membrane 39

III. Inorganic Ions, Physical Chemistry, and Cell Physiology 39 1. Bicarbonate, Carbon Dioxide, pH, and Buffering . 40 2. Sodium, Chloride, Osmolarity, and Humidification of

Incubators 41 3. Potassium and Na + : K + Ratios 42 4. Calcium, Magnesium, and Regulatory Roles of Divalent Cations 43 5. Phosphate. 44 6. Other Physicochemical Parameters 45

IV. Qualitative Nutrient Requirements 46 1. Components of Eagle's Minimum Essential Medium 46 2. Other Amino Acids 47 3. Other Vitamins 47 4. Carbohydrates and Intermediates of Energy Metabolism. 48 5. Nucleic Acid Components. 49 6. Other Organic Nutrients 50 7. Inorganic Trace Elements 50

V. Quantitative Optimization of Synthetic Media 52 1. Species and Cell Type Individuality of Quantitative

Requirements 52 2. Reduction of Requirements for Serum and Growth Factors 53

VI. Lipids and Related Substances 54 1. Fatty Acids 54 2. Prostaglandins . 55 3. Phospholipids 56 4. Cholesterol 56 5. Synthetic Media 56

VII. Hormones, Hormone-Like Growth Factors, and Carrier Proteins 57 1. Hormones. 57 2. Growth Factors 57 3. Replacement of Serum with Hormones and Growth Factors 59

Contents XI

4. Individuality of Cellular Requirements 59 5. Nontransformed Cells . . . . . . . 60 6. Relationship to In Vivo Growth Requirements 61

VIII. Special Requirements Related to Cellular Density 62 1. Requirements for Clonal Growth. . . . . . 62 2. Requirements for Multiplication of Dense Cultures 63

IX. Regulatory Interactions and Artificial Stimulation of Multiplication . . . . 64 1. Borderline Toxicity. 65 2. Proteolytic Enzymes 65 3. Tumor Promoters 65 4. Inorganic Ions. . . 66 5. Cyclic Nucleotides . 66 6. Defining a Genuine Growth Requirement 67

D. Requirements for Survival of Nontransformed Cells Without Proliferation. . . . . 67

I. Early Studies. . . . . . . . . . . . . . 68 II. Survival Factor. . . . . . . . . . . . . 68

III. Elimination of the Need for Survival Factor 68 IV. Nutrients and Survival . . . . . . . . . 69 V. Differences in Survival Between Nontransformed and Transformed

Cells . . . . . . . . . . . . . . . . . . . . . . 69 E. Future Studies of the Growth and Survival Requirements of

Nontransformed Cells . . . . . . . . . . . . . . . . . 70 I. Integration of New Findings . . . . . . . . . . . . 70

II. Questions Remaining to be Solved After Development Synthetic Media. . . . . . . . . . . . 70

III. Magnitude of Remaining Work. 71 IV. Selective Allocation of Resources 72

F. Summary and Conclusions 72 G. Note Added in Proof. 73 References . . . . . . . . . 74

CHAPTER 3

Epidermal Growth Factor. G. CARPENTER. With 3 Figures

A. Introduction. . . . . . . . . . . . . B. Chemical and Physical properties of EGF

I. Mouse EGF. . . . . . . . . . . 1. Isolation . . . . . . . . . . . 2. Chemical and Physical Properties 3. Derivatives

a) EGF-2 ....... . b) EGF-5 ....... . c) Cyanogen Bromide EGF .

4. High Molecular Weight Mouse EGF .

89 89 89 89 90 91 91 91 92 92

XII

II. Human EGF . . . . . . . . . . 1. Identification and Isolation . . . 2. Chemical and Physical Properties 3. Relationship of Human EGF and Urogastrone

III. Rat EGF . . . . . . . . .

C. Physiological Aspects of EGF . . 1. Concentration in Body Fluids

1. Mouse EGF 2. Human EGF

II. Localization. . 1. Mouse EGF 2. Human EGF

III. Control of Submaxillary Gland Content of Mouse EGF . IV. Secretion of Mouse EGF from the Submaxillary Gland V. Factors Affecting Levels of Human EGF

D. Biological Activities of EGF In Vivo 1. Skin . . . . . ...

II. Corneal Epithelium III. Respiratory Epithelium IV. Gastrointestinal Tract V. Liver ....... .

E. Organ Culture Studies of EGF . 1. Skin . . . .

II. Other Tissues 1. Cornea. 2. Palate .. 3. Bone

F. Cell Culture Studies of EGF . 1. Cell Nutrition . . . . .

II. Types of Cells Affected by EGF III. EGF and the Growth of Cell Populations IV. Components of the Mitogenic Response

1. Rapid Biological Responses at the Membrane a) Uridine Uptake b) Sugar Transport . . c) Cation Fluxes . . . d) Putrescine Transport e) Alanine Transport . f) Membrane Ruffling and Macropinocytosis g) Other Membrane Responses . . . . . .

2. Responses of EG F Occurring in the Cytoplasm a) Activation of Glycolysis . . . . . . . . b) Synthesis of Extracellular Macromolecules c) Activation of RNA and Protein Synthesis d) Activation of Ornithine Decarboxylase. e) Protein Phosphorylation. . . . . . . .

Contents

93 93 93 94 95 95 95 95 95 95 95 96 96 97 97 98 98 99 99 99

100

100 100 101 101 102 102 102 102 103 103 104 104 104 105 105 105 106 106 106 106 107 107 107 108 108

Contents XIII

3. Stimulation of DNA Synthesis 108 V. Responses Not Related to Mitogenesis 109

G. Growth Factor: Receptor Interactions. . . 109 I. Receptors for EGF. . . . . . . . . 109

II. Internalization and Degradation of EGF 110 1. Biochemical Evidence . . . 110 2. Morphological Evidence . . 111

III. Internalization of the Receptor. 112 1. Indirect Evidence . . . 112 2. Chemical Evidence. . . . . 113 3. Morphological Evidence . . 113

IV. Recovery of Receptor Activity. 114 H. Relationship of EGF Binding and Metabolism to Biological Activity 115

1. Rapid Changes in Cell Physiology 115 II. Stimulation of DNA Synthesis. 116

I. Other Controls of Receptor Activity 118 I. Transforming Agents . 118

II. Tumor Promoters . . . . . . 119 III. Differentiation. . . . . . . . 120 IV. Lectins and Glycoprotein Metabolism. 120 V. Glucocorticoids . . . . . . . . . . 121

VI. Modulation of Protein Synthesis . . . 121 K. A Biochemical Response to EGF in Subcellular Systems. 121 L. Prospectus 122 References . . . . . . . . . . . . . . . . . . . . . . 123

CHAPTER 4

The Platelet-Derived Growth Factor. R. Ross. With 14 Figures

A. Serum, the Platelet-Derived Growth Factor, and Cell Culture 133 B. The Platelet. . . . . . . . . 135 C. Platelet-Structure and Function . . . . . . . . . . . . . 136 D. The Gray Platelet Syndrome . . . . . . . . . . . . . . 138 E. The Megakaryocyte as the Source of Platelet-Derived Growth Factor. 138 F. Purification and Characterization of the Platelet-Derived Growth Factor 140 G. The Spectrum of Cell Response . . . . . . . . . . . . . . 142 H. Control of Cell Proliferation by Platelet-Derived Growth Factor

and Plasma . . . . . . . . . . . . . . . . . . 144 I. The Role of Plasma . . . . . . . . . . . . . . . . . 146

K. Endocytosis and the Platelet-Derived Growth Factor . . . 149 L. Modulation of Receptors for Epidermal Growth Factor by

Platelet-Derived Growth Factor . . . . . . . . . . . . 149 M. Lipid Metabolism and the Platelet-Derived Growth Factor 150 N. Platelets and Cell Proliferation In Vivo 154 O. Summary. 156

References . . . . . . . . . . . . . . 157

XIV Contents

CHAPTER 5

Somatomedin: Physiological Control and Effects on Cell Proliferation D. R. CLEMMONS and J. J. VANWYK. With 13 Figures

A. Introduction. . . . . . . . . . . . . . . . 161 B. Assay Systems Used to Measure Somatomedins 162

I. Biological Assays 162 II. Radioreceptor Assays. 163

III. Protein Binding Assays 163 IV. Radioimmunoassays . 163 V. Standards Used for the Quantitation of Somatomedin Activity 166

C. Isolation and Properties of the Individual Somatomedins 166 I. Basic Somatomedins . . . . . . . . . 166

1. Somatomedin-C (SM-C) . . . . . . 166 2. Insulin-Like Growth Factor I (IGF-I) 169 3. Somatomedin in Other Species 170

II. Neutral Somatomedins . . . . . . . . 171 1. Insulin-Like Growth Factor II (IGF-II) 171 2. Somatomedin-A (SM-A) . . . . . . . 171 3. Multiplication Stimulating Activity (MSA) 171

D. Production of the Somatomedins. . . . . . . . 172 I. Somatomedin Production by Organs and Tissue Slices. 172

II. Somatomedin Production by Monolayer Cultures. . . 174

E. Molecular Size and Transport of Somatomedins in Plasma: The Somatomedin Binding Proteins. . . . . . . 175

F. Control of Somatomedin Concentrations in Blood 178 I. Blood Concentrations in Normal Individuals. 178

1. Effect of Age . . . . . . 178 2. Effect of Hormonal Status . . . . . 180

a) Growth Hormone. . . . . . . . 180 b) Prolactin and Placentral Lactogens 182 c) Thyroid Hormone 183 d) Cortisol . . . . 183 e) Estrogens . . . . 183

3. Effect of Pregnancy . 184 4. Effect of Nutritional Status 184

G. In Vitro Biological Effects of the Somatomedins 186 I. Whole Tissue Effects 186

1. Cartilage . . . 186 2. Muscle. . . . . 187 3. Adipose Tissue . 187

II. Correlation Between Biological Responses and Receptor Interactions 188 III. Stimulation of DNA Synthesis and Cell Growth in Tissue Culture 190

1. Range of Responsive Cell Types. . . . . . . . . . . . . . 190 2. Interaction Between Somatomedin and Other Growth Factors

in the Cell Cycle. . . . . . . . . . . . . . . . . . . . . 191

Contents XV

3. Production of Somatomedin-Like Peptides by Cultured Cells and Their Role in Cellular Proliferation 193

H. In Vivo Actions of the Somatomedins 196 References . . . . . . . . . . . . . . . . . . 198

CHAPTER 6

Glucocorticoid Modulation of Cell Proliferation V. J. CRISTOFALO and B. A. ROSNER. With 2 Figures

A. Introduction. . . . . . . . . . . . . . . . I. Historical Perspective. . . . . . . . . .

II. Glucocorticoids: General Mechanisms of Action B. Hormone Responsiveness by Cell Cultures.

I. Various Vertebrate Species II. Human Cell Lines . . . . . .

III. Mouse and Rat Cell Lines C. Permissive Effects of Glucocorticoids

I. Glucocorticoid Modulation of Heterologous Receptors II. Glucocorticoid Effects on Other Factors Implicated in the

Regulation of Cell Growth . . . . . . . . . . . . . . D. Glucocorticoid Induction of Growth Factors in Normal Human Cells

In Vitro E. Summary and Conclusions References . . . . . . . . .

CHAPTER 7

Proteases as Growth Factors. D. D. CUNNINGHAM

209 209 209 211 211 213 215 218 218

219

220 222 224

A. Introduction. . . . . . . . . . . . . . . . . . . . . . . .. 229 B. Stimulation of DNA Synthesis and Cell Division by Proteases . .. 229

I. Cell-Protease Combinations Which Yield Mitogenic Stimulation 229 II. Involvement of Proteases in the Action of Other Growth Factors 231

III. Protease Activity of Other Growth Factors . . . . . . . . . . 232 C. Mechanisms of Protease-Stimulated Cell Proliferation. . . . . . . . 233

1. Mitogenic Stimulation by Proteases Under Serum-Free Chemically Defined Conditions ....... 233

II. Requirement of Proteolytic Activity. 234 III. Sufficiency of Cell Surface Action . 235 IV. Cell Surface Receptors for Thrombin 236 V. Cleavage of Cell Surface Proteins and Stimulation of Cell Division 238

VI. Perspectives and Future Studies . . . . . . . . . . . . . . . 239 D. Possible Role of Protease-Stimulated Cell Proliferation in Physiological

Processes . 240 E. Conclusions 242 References . . 243

XVI Contents

CHAPTER 8

Nerve Growth Factor. MARY J. KOROLY and M. YOUNG

A. Discovery of NGF and Its Early History B. Multiple Molecular Forms of NGF .

I. Cohen's NGF II. 7S NGF .....

III. 2.5S NGF. . . . . IV. Snake Venom NGF V. What is the Naturally Occurring Form of NGF in the Mouse

249 250 250

· 251 252

· 253

Salivary Gland? . . . . . . . . . . . . . 254 C. The NGF-Zymogen and Its Enzymic Properties . 255

I. Autocatalytic Self-activation. . . . . . . . 255 II. Regulation of the Autoactivation Reaction . 257

III. Activation of Plasminogen by NGF. . . . . 257 D. Role of the Submandibular Gland in Secretion of NGF . 258 E. Secretion of NGF by Cells in Culture. . . . . . . . . 258 F. Biological Effects of NGF Related to the Nervous System and Neural

Crest Derivatives. . . . . . . . . . . 260 I. Sympathetic and Sensory Neurons . 260

II. Adrenal Cells . . . . . . 263 III. Pheochromocytoma Cells . . . . . 263 IV. Central Nervous System . . . . . 264

G. Retrograde Transport and Trophic Effects . 265 H. Cellular NGF Receptors . . . . . . . . . 265 I. Other Biological Actions of the 116000 Molecular Weight NGF . 266

I. Effect of Saliva and NGF on Wounds . . . 267 II. Effect of Saliva and NGF on Stress Ulcers . 267

K. Summary and Perspectives . 267 References . . . . . . . . . . . . . . . . . . . 268

CHAPTER 9

The Role of Cold Insoluble Globulin (Plasma Fibronectin) in Cell Adhesion In Vitro. F. GRINNELL. With 9 Figures

A. Introduction. . . . . . . . . . . . . . . . . . . . . . 277 B. Cold Insoluble Globulin and Fibronectin: Terminology . . . 278 C. The Role of CIG in Cell Adhesion to Tissue Culture Dishes . 278

I. Historical Overview . . . . . . . . . . . . . . . . 278 II. Identification of CIG as the Serum Factor in Cell Attachment and

Spreading. . . . . . . . . . . . . . . . . . . . 279 III. Analysis of CIG Adsorption to Tissue Culture Dishes. 280 IV. Technical Comments on CIG Purification . . . . . . 282 V. Other Components Which Promote Cell Spreading . . 284

D. The Role of CIG in Cell Adhesion to Surfaces Other than Tissue Culture Dishes. . . . . . · 286

I. Bacteriological Dishes . . . . . . . . . . . . . . . . . · 286

Contents XVII

II. CIG and the Adhesion of Cells to Collagen . . . . . . . 286 III. CIG and the Adhesion of Cells to Fibrin and Fibrinogen 290

E. Cell Spreading in the Absence of Serum or CIG 290 F. Mechanism of Action of CIG . . . 291

I. Ligand Receptor Hypothesis ...... 291 II. Cooperativity and CIG Binding . . . . . 292

III. Interaction of Multimeric CIG with BHK Cells 293 IV. Inhibition of Cell Adhesion with Gangliosides 294 V. Isolation of Cell-Substratum Adhesion Sites 294

VI. Studies on the Active Site(s) of CIG 294 G. Summary 295 References . . . . . . . . . . . . . . . 295

CHAPTER 10

Membrane-Derived Inhibitory Factors. P. DATTA. With 4 Figures

A. Introduction and Perspective ................. 301 B. Some Properties of Membrane-Derived Inhibitory Factors. . . .. 303 C. Cell Cycle-Dependent Inhibition of DNA Synthesis and Cell Division 304 D. The Role of Glycosylation in the Control of DNA Synthesis . 306 E. Inhibition of DNA Synthesis in Virus-Transformed Cells 308 F. Surface Membranes and Nutrient Uptake 309 G. Prospects and Conclusions 3lO References . . . . . . . . . . . . . . . 311

CHAPTER II

Diffusible Factors in Tissue Cultures. LOUISE HAREL

A. Introduction. . . . . . . . . B. Metabolic Cooperation . . . . C. Diffusion of Stimulating Factors

I. The Feeder Effect . . . . II. Multiplication-Stimulating Activity from Rat Cells

1. Purification of MSA . . . 2. Metabolic Effects of MSA 3. Cell Surface Receptors . .

D. Diffusion of Inhibitory Factors I. Problem of Density-Dependent Inhibition

1. Mechanism of DDI . . . . . . II. Inhibitory Factors Released by Cells

I. Inhibitors from 3T3 Cells. . . . 2. Inhibitors from WI38 Cells . . . 3. Inhibitors from Chinese Hamster Fibroblasts 4. Inhibitors from a Melanocytic Line 5. Inhibitors from BSC1 Cells . . . . . . . .

313 314 318 318 320 320 321 322 322 322 323 326 326 329 329 330 330

XVIII Contents

E. Diffusion of Transforming Factors . . . . . . . . . . . . I. Diffusion of Protein Factors Which Enhance Malignant

Transformation . . . . . . . . . . . . . . . . II. Metabolic Interaction Which Enhances the Chemical

Transformation of Cells F. Concluding Remarks References . . . . . . . . . .

CHAPTER 12

Hemopoietic Colony Stimulating Factors. D. METCALF With 6 Figures

A. Introduction and Terminology. . . . . . . . . B. Culture of Hemopoietic Colonies in Semisolid Medium C. Detection and Assay of Colony-Stimulating Factors D. Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF)

I. Sites of Production of GM-CSF . . . . . . . II. Purification and Chemical Nature of GM-CSF .

III. Mechanisms of Action of GM-CSF ..... . IV. Factors Modifying Responsiveness to GM-CSF V. Factors Influencing GM-CSF Production and Levels

1. Steady State Production of GM-CSF ..... . 2. Increased GM-CSF Production in Response to Infections and

Bacterial Products. . . . . . . . . . . . . . . . . 3. Increased GM-CSF Production Following Lymphocyte

Stimulation. . . . . . . . . . . . . . . 4. Other Situations Modifying GM-CSF Levels

VI. Role of GM-CSF In Vivo ......... . E. Eosinophil Colony Stimulating Factor (EO-CSF). . F. Megakaryocyte Colony Stimulating Factor (MEG-CSF) . G. Erythropoietin and Erythroid Colony Stimulating Factor H. Final Comments References .

CHAPTER 13

331

331

332 333 334

343 344 345 347 347 350 353 359 360 361

361

364 365 367 369 370

· 371 374

· 374

Inhibition of Hematopoietic Cell Proliferation. J. H. FITCHEN and M. J. CLINE With 3 Figures

A. Introduction. . . . . . . . . . . . . . . B. Hematopoietic Techniques and Nomenclature

I. Pluripotent Stem Cells . . . . . II. Unipotent (Committed) Stem Cells . . .

C. Inhibitors of Hematopoiesis . . . . . . . . I. "Physiologic" Inhibitors of Hematopoiesis

1. Prostaglandins . . . . . . . . . . . 2. Neutrophil-Derived Colony Inhibitory Factor 3. Chalones. . . . 4. Serum Inhibitors . . . . . . . . . . . .

385 387 387 389 389 390 391 392 392

· 393

Contents

II. Inhibitors of Hematopoiesis in Disease 1. Leukemic Inhibitors . . . . . . . 2. Immune Suppression of Hematopoiesis .

a) Humoral Inhibitors . . . . . . . . b) Cell-Mediated Immune Suppression

D. Summary and Conclusions References . . . . . . . . . . . . . . . . .

CHAPTER 14

Inducers and Inhibitors of Leukemic Cell Differentiation in Culture JANET ABRAHM and G. ROVERA

A. Introduction. . . . . . . . . . . . . . . . . . . . . . B. Leukemic Cell Lines Utilized for Differentiation Studies in Culture C. Inducers of Differentiation in Culture. . . . . . . D. Mechanism of Action of Inducers of Differentiation . . . . E. Clonal Analysis of the Induction of Differentiation . . . . . F. Inducers of Leukemic Cell Differentiation and the Cell Cycle G. Inhibitors of Differentiation . . . . . . . . . . . . . . . H. Inducers of Differentiation and Treatment of Leukemias In Vivo References .

CHAPTER 15

Angiogenesis Factor(s). P. M. GULLINO. With 2 Figures

XIX

393 393 394 395 398 400

. 400

405 406 407 413 415 416 416 418

. 420

A. Introduction. . . . . . . . . . . . 427 B. Morphogenesis of Vascular Networks . 427 C. Assays for Angiogenesis. . . . . . . 429

I. Rabbit Eye . . . . . . . . . . 429 II. Chick Embryo Chorioallantoic Membrane. 431

III. Hamster Cheek Pouch . . . . . . . . . 432 IV. Dorsal Air Sac and Intracutaneous Injections 433 V. Endothelial Cell Culture 433

VI. Renal Assay. . . . . . . . . 434 D. Induction of Angiogenesis. . . . . . 434

I. Angiogenesis by Normal Tissues 434 II. Angiogenesis by Normal Cells . 436

III. Angiogenesis by Neoplastic Tissues and Cells 437 E. Isolation and Characterization of Angiogenesis Factor(s) 438

I. Fractionation of Fluids or Tissues with Angiogenic Capacity . 439 II. Angiogenic Capacity of Growth Factors and Prostaglandins 440

F. Physiologic Significance of Angiogenesis Factor(s) . . . . . . 441 I. Angiogenesis as a Marker for Neoplastic Transformation . 442

II. Antiangiogenesis . . 443 G. Concluding Remarks . 444 References . . . . . . . . 445

xx Contents

CHAPTER 16

Growth of Human Tumors in Culture. HELENE S. SMITH and CH. M. DOLLBAUM. With 1 Figure

A. General Introduction . . . . . . . . . . . . . . . I. The Ideal System .. . . . . . . . . . . . .

II. Current State of Technology for Culturing Tumor Cells III. Identifying Cell Types in Culture.

1. Endothelial Cells . 2. Epithelial Cells . . . . . . . 3. Fibroblastoid Cells ... . .

IV. Identifying Tumor Vs Nonmalignant Cells in Culture V. Identifying Cell-to-Cell Contamination in Culture

B. Specific Systems . . . . I. Carcinomas . . . . .

1. Mammary Gland . 2. Bladder and Kidney 3. Colon ..... 4. Prostate . . . .

II. Other Malignancies 1. Melanomas . 2. Gliomas . 3. Sarcomas

C. Summary References .

CHAPTER 17

The Chalones. O. H. IVERSEN. With 4 Figures

451 451 454 456 457 458 459 460 463 464 465 465 469 470 471 473 473 476 477 478

. 478

A. Introduction. . . . . . . . . . 491 I. Definition and Properties . 491

II. Limitations of This Chapter 492 III. The Name. . . . . . . . 493 IV. Theoretical and Biological Background 493

1. Theory . . . . . . . . . . . . . 493 2. Biology. . . . . . . . . . . . . 494

B. Sources of Chalones: Methods to Extract, Purify and Measure Their Effects . . . . . . . . . . . . . . . . . . . . . 495

I. Sources of Chalones. . . . . . . . . . . . . . . . 495 II. Methods to Assess Chalone-Mediated Growth Inhibition. 496

III. Methods to Purify and Characterize Chalones 497 C. Some Chalone Properties . . . . . . . . . . . 497

I. Tissue or Cell Line Specificity or Preference 497 II. Species Non-specificity . . . . . . . . . 499

III. Sites of Attack in the Cell Cycle . . . . . 499 IV. Reversibility and Turnover Time of Chalones 500 V. Chalones and Stimulators of Cell Proliferation 501

Contents XXI

VI. Dose-Response Relationship 501 VII. Chemical Composition 502

D. Mechanisms of Action . . . . . 502 I. General Considerations . . 502

II. Do Chalones Primarily Inhibit Proliferation, or Do They Promote Maturation? . . . . . . . . . . . . . . . . . . 503

III. Possible Relationship to Hormones and Cyclic AMP. 503 IV. Do Chalones Act via the Cell Membranes? . 504

E. Chalones and Malignancy. . . . . . 504 I. General Considerations . . . . 504

II. Chalones in Malignant Tumors . 505 III. Chalones and Carcinogenesis. 505

F. Possible Practical Uses of Chalones .' 506 I. For Diagnostic Purposes. . . 506

II. For Therapeutic Purposes . . 506 1. Diseases with Benign Increased Cell Proliferation . 506 2. Treatment of Cancer . . . 506 3. Immunosuppression 509 4. A Male Antifertility Drug . 509

G. The Various Chalones . . . . 509 I. The Epidermal Chalones. . . 509

1. The Epidermis. . . . . . 509 2. The Epidermal G2 Chalone 509 3. The Epidermal G 1 Chalone 511 4. Conclusions About Epidermal Chalones . 512

II. Chalones from Epidermal Derivatives . . . 512 1. The Mammary Gland Chalone? . . . . 512 2. Sebaceous Gland, Sweat Gland and Hair Follicle Chalones? 513

a) Sebaceous Gland . . . . . . 513 b) Sweat Gland (Eccrine) Chalone 513 c) Hair Root Chalone . . . . . 513

III. The Melanocyte Chalone (?) . . . . 514 IV. Chalones in the Cells of the Red Bone Marrow, Blood and

Lymphoid System. . . . . 514 1. The Granulocyte Chalone. . 516 2. The Erythrocyte Chalone . . 518 3. The Lymphocyte Chalone(s) . 519 4. The Monocyte (Macrophage) Chalone (?) 523 5. The Platelet Chalone (?). . . . . . . 523 6. The Stem Cell Chalone (?). . . . . . 523 7. Conclusion About Blood Cell Chalones 524

V. Chalones in the Gastro-intestinal Tract . 524 1. Salivary Gland Chalone (?) . . . . . 524 2. Oral Mucosa, Oesophageal and Forestomach Chalone 524 3. Gastric Chalone (?). . . 524 4. Small Intestinal Chalone 524 5. A Colon Chalone? . . . 525

XXII

VI. The Liver Chalones . . . . . . . . . . . . VII. Kidney Chalones (?) ........... .

VIII. Chalones in the Male Reproductive Organs (?) 1. Testicular Chalone . . . . . . . 2. A Chalone in the Seminal Vesicle? 3. A Prostatic Chalone? . . . . . .

IX. Ascites Tumour Cell Chalones . . . 1. JBI Ascites Tumour Cell Chalone 2. Ehrlich Ascites Tumour Cell Chalone .

X. Connective Tissue and Fibroblast Chalones (?) XI. Chalones of the Lung?

XII. Other Chalones Indicated 1. Lens of the Eye . . . 2. Endothelial Chalone? . 3. Smooth Muscle Chalone? 4. Heart Muscle Chalone? 5. Placental Chalone?

References Addendum ........ .

Contents

525 526 527 527 528 528 529 529 529 530 531 531 531 531 531 531 532 532 549

Author Index . . . . . . . . . . . . . . . . . . . ........ 551

Subject Index ........................... 627