Koller / Palsson / Masters Human Cell Culture

Primary Hematopoietic Cells
1. Auflage 2006
ISBN: 978-0-306-46886-5
Verlag: Springer Netherlands
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 4, 352 Seiten

Reihe: Human Cell Culture

Cell Biology - Short description currently not available.

Autoren/Hrsg.

Koller, F.

Palsson, Bernhard Ø

Palsson, B.

Masters, John

Masters, J.R.

(Eds.), UK

Inhaltsverzeichnis

1;Table of Contents;6
2;Introduction;7
3;Hematopoietic Stem and Progenitor Cells;8
3.1;1. INTRODUCTION;8
3.1.1;1.1 Function and Organization of the Hematopoietic System;8
3.1.2;1.2 Stem Cell Self-Renewal;9
3.1.3;1.3 Hematopoietic Cell Growth Factors;11
3.1.4;1.4 The Bone Marrow Microenvironment;11
3.2;2. TISSUE PROCUREMENT AND PROCESSING;13
3.2.1;2.1 Human Hematopoietic Tissue Procurement and Shipping;13
3.2.2;2.2 Human Hematopoietic Tissue Processing;15
3.3;3. ASSAYS;16
3.3.1;3.1 Non-Biological Assays;16
3.3.2;3.2 Biological Assays;18
3.4;4. CULTURE TECHNIQUES;20
3.4.1;4.1 Historical Perspective: Long-Term Bone Marrow Culture ( LTBMC);20
3.4.2;4.2 Current Culture Techniques;21
3.5;5. UTILITY OF HEMATOPOIETIC STEM AND PROGENITOR CELL CULTURES 5.1 Clinical Background;26
3.5.1;5.1 Clinical Background;26
3.5.2;5.2 Clinical Use of Hematopoietic Cell Cultures;27
3.5.3;5.3 Other Uses of Hematopoietic Cell Cultures;28
3.6;6. SUMMARY;28
3.7;LITERATURE CITED;29
4;In Vitro T-Lymphopoiesis;38
4.1;1. INTRODUCTION;38
4.2;2. PROTOCOLS FOR IN VITRO T-CELL DIFFERENTIATION;41
4.2.1;2.1 T-cell Differentiation Using Rhesus Thymic Stroma ;41
4.2.2;2.2 T-cell Differentiation Using Human Thymic Stroma;43
4.2.3;2.3 Thymic Organ Culture;44
4.2.4;2.4 Reconstituted Human-SCID Mice Thymic Cultures;45
4.3;3. ANALYSIS OF CELL OUTPUT 3.1 Immunophenotyping of Cultured Cells by Flow Cytometry;45
4.3.1;3.1 Immunophenotyping of Cultured Cells by Flow Cytometry;45
4.3.2;3.2 RT-PCR Analysis;46
4.4;4. UTILITY OF THE SYSTEM;48
4.5;5. CONCLUDING REMARKS;49
5;T-Lymphocytes: Mature Polyclonal and Antigen- Specific Cell Culture;52
5.1;1. INTRODUCTION;52
5.1.1;1.1 Anatomy In Vivo;53
5.1.2;1.2 T-cell Activation;54
5.1.3;1.3 The Beginnings of T-Lymphocyte In Vitro Culture;57
5.2;2. TISSUE PROCUREMENT AND PROCESSING;57
5.2.1;2.1 Procurement;57
5.2.2;2.2 Enrichment/Purification from Whole Blood or an Apheresis Product;59
5.2.3;2.3 Selection of T-Cells;61
5.2.4;2.4 Cryopreservation;64
5.3;3. CULTURE TECHNIQUES;64
5.3.1;3.1 Media and Serum;65
5.3.2;3.2 Culture Vessels and Large-Scale Culture;66
5.3.3;3.3 Polyclonal Activation and Culture of T-Cells;68
5.3.4;3.4 Development of Ag-Specific T-cell Lines;75
5.4;4. ASSAY TECHNIQUES;81
5.4.1;4.1 Proliferation;81
5.4.2;4.2 Tracking Cell Division in T-cell Subsets;84
5.4.3;4.3 TCR Repertoire;86
5.4.4;4.4 Flow Cytometry and T-cell Culture;87
5.4.5;4.5 Cytokine Secretion and Intracellular Cytokine Flow Cytometry;88
5.4.6;4.6 CTL Activity;89
5.5;5. UTILITY OF THE SYSTEM;90
5.6;6. SUMMARY;92
5.7;ACKNOWLEDGEMENTS;93
5.8;REFERENCES;93
6;The Culture, Characterization and Triggering of BLymphocytes;108
6.1;1. INTRODUCTION;108
6.1.1;1.1 Origin of Normal B- Lymphocytes;109
6.1.2;1.2 Historical Aspects of the Culture of B- Lymphocytes;109
6.2;2. TISSUE PROCUREMENT AND PROCESSING ;111
6.2.1;2.1 Sources of Normal B-Lymphocytes;111
6.2.2;2.2 Separation of Peripheral Blood and Bone Marrow BLymphocytes;111
6.2.3;2.3 Separation of Splenic and Tonsillar B- Lymphocytes;112
6.2.4;2.4 Purification of B-Lymphocytes;113
6.3;3. ASSAY TECHNIQUES;115
6.3.1;3.1 B-Lymphocyte Antigens;115
6.3.2;3.2 Assays of B-Lineage Active Cytokines;116
6.4;4. CULTURE TECHNIQUES ;117
6.4.1;4.1 Media and Conditions;117
6.4.2;4.2 Influence and Dependence on Growth Factors;118
6.4.3;4.3 Long - Term Culture Using Adherent Feeder Cell Layers;118
6.4.4;4.4 NIH3T3 CD40-Ligand Transfectants;123
6.5;5. UTILITY OF SYSTEM;125
6.5.1;5.1 B-Cell Differentiation;125
6.5.2;5.2 B- Cell Growth;125
6.5.3;5.3 B-Cell Signaling;126
6.5.4;5.4 B-Cell Survival and Apoptosis;126
6.6;6. CONCLUSION;126
6.7;LITERATURE CITED;127
7;Monocytes and Macrophages;132
7.1;1. INTRODUCTION;132
7.2;2. TISSUE PROCUREMENT AND PROCESSING;134
7.2.1;2.1 The Cell Source Limits the Type of Investigation;137
7.2.2;2.2 To Purify or Not to Purify;137
7.2.3;2.3 To Freeze or Not to Freeze;139
7.3;3. ASSAY TECHNIQUES ;139
7.3.1;3.1 General Characteristics;139
7.3.2;3.2 Flow Cytometry;140
7.3.3;3.3 In Vitro Production of Macrophages and Detection of Monocyte-Macrophage Progenitor Cells Using the Colony-Forming Technique;140
7.4;4. CULTURE TECHNIQUES;145
7.4.1;4.1. Type of Growth Surface;145
7.4.2;4.2 Serum or Serum-Free;146
7.4.3;4.3 Other Culture Components;146
7.4.4;4.4 Growth Factors;146
7.4.5;4.5 Low Oxygen Tension;147
7.4.6;4.6 Production of Resident Macrophages on a Nonadherent Growth Surface;147
7.5;5. UTILITY OF THE SYSTEM;148
7.6;6. CONCLUDING REMARKS;149
7.7;ACKNOWLEGMENT;149
7.8;REFERENCES CITED;149
8;Isolation and Cultivation of Osteoclasts and Osteoclast-Like Cells;153
8.1;1. INTRODUCTION;153
8.2;2. TISSUE PROCUREMENT AND PROCESSING;156
8.2.1;2.1 Human Osteoclasts (In Vivo Formed);156
8.2.2;2.2 Human Osteoclast-Like Cells (In Vitro Formed);159
8.3;3. CULTURE TECHNIQUES 3.1 Preparation of Sterile Devitalized Bone or Ivory Discs for Resorption Studies;160
8.3.1;3.1 Preparation of Sterile Devitalized Bone or Ivory Discs for Resorption Studies;160
8.3.2;3.2 Culture of In Vivo Formed hOC;161
8.3.3;3.3 Culture of In Vitro Formed hOCL;162
8.4;4. ASSAY TECHNIQUES 4.1 Morphology and Ultrastructure;163
8.4.1;4.1 Morphology and Ultrastructure;163
8.4.2;4.2 Cytochemical Staining;164
8.4.3;4.3 Antigenic Profile;166
8.4.4;4.4 Molecular Profile;168
8.4.5;4.5 Bone Resorptive Function;168
8.5;5. UTILITY OF SYSTEM;170
8.6;6. CONCLUDING REMARKS;171
8.7;ACKNOWLEDGEMENTS;172
8.8;LITERATURE CITED;172
9;Isolation and Culture of Dendritic Cells;176
9.1;1. INTRODUCTION;176
9.2;2. ISOLATION OF DENDRITIC CELLS;178
9.2.1;2.1 Isolation of Specific Types of Dendritic Cells: Langerhans Cells;181
9.2.2;2.2 Other Methods to Increase the Number of Isolated Dendritic Cells;181
9.3;3. GENERATION OF DENDRITIC CELLS;182
9.3.1;3.1 Generation of DC from CD34+Cells;183
9.3.2;3.2 Generation of DC from CD14+ and Plastic-Adherent Cells;184
9.3.3;3.3 Choice of Medium;186
9.3.4;3.4 Methods for Loading DC with Antigen;188
9.4;4. ASSAYS FOR ASSESSING DENDRITIC CELLS;189
9.5;5. FUTURE DIRECTIONS;190
9.6;LITERATURE CITED;191
10;In Vitro Proliferation and Differentiation of CD34+ Cells to Neutrophils;197
10.1;1. INTRODUCTION;197
10.2;2. TISSUE PROCUREMENT AND PROCESSING ;198
10.2.1;2.1 Obtaining CD34+ Cells;198
10.2.2;2.2 Preparation of Mat;199
10.2.3;2.3 Maintainance of Mature Cells In Vitro;199
10.3;3. ASSAY TECHNIQUES;200
10.3.1;3.1 CD34+ Cells;200
10.3.2;3.2 Measurement of Neutrophil Differentiation and Function;200
10.4;4. CULTURE TECHNIQUES ;206
10.4.1;4.1 System Concepts;206
10.4.2;4.2 Static Systems: Gas Permeable Bags;206
10.4.3;4.3 Cell Culture Bag Materials;207
10.4.4;4.4 Optimal Volumes and Cell to Surface Area Ratio;207
10.4.5;4.5 Perfusion Systems;208
10.4.6;4.6 Growth Factors and Medium;209
10.4.7;4.7 Characterization of In Vitro Derived Neutrophils;210
10.5;5. UTILITY OF SYSTEM;212
10.6;6. CONCLUDING REMARKS;215
10.7;7. LITERATURE CITED;216
11;Isolation and Culture of Eosinophils;222
11.1;1. INTRODUCTION;222
11.2;2. ISOLATION OF EOSINOPHILS AND EOSINOPHIL PROGENITORS ;224
11.2.1;2.1 Normal Peripheral Blood Eosinophils;224
11.2.2;2.2 Hypereosinophilic Donors;227
11.2.3;2.3 Umbilical Cord Blood Progenitors;227
11.2.4;2.4 Peripheral Blood and Bone Marrow Progenitors;229
11.3;3. IDENTIFICATION OF MATURE EOSINOPHILS;229
11.3.1;3.1 Morphology and Staining Characteristics;229
11.3.2;3.2 Cell Surface Markers;229
11.3.3;3.3 Specific Granule Proteins;230
11.4;4. METHODS FOR EOSINOPHIL CULTURE ;230
11.4.1;4.1 Maintaining Viability of Eosinophil Isolated from Pheripheral Blood;230
11.4.2;4.2 Differentiation and Culture of Eosinophils from BM and CB Progenitors;231
11.4.3;4.3 Differentiation and Culture of Eosinophils from Peripheral Blood Progenitors;232
11.4.4;4.4 Quantitative Comparisons Between Natural and Cultured Eosinophils;232
11.5;5. APPLICATIONS ;234
11.5.1;5.1 Characterization of Eosinophil-Mediated Functions In Vitro;234
11.5.2;5.2 Apoptosis and the Control of Eosinophilic Inflammation;234
11.5.3;5.3 Molecular Mechanisms of Differentiation and Transcriptional Control of Eosinophil- Specific Genes;235
11.5.4;5.4 Identification of Novel Enhancers and Inhibitors of Eosinophil Differentiation;235
11.6;REFERENCES;235
12;Isolation and Culture of Mast Cells and Basophils;243
12.1;1. INTRODUCTION;243
12.2;2. TISSUE PROCUREMENT AND PROCESSING ;246
12.2.1;2.1 Progenitor Cells;246
12.2.2;2.2 Peripheral Blood Basophils;247
12.2.3;2.3 Tissue Mast Cells;248
12.2.4;2.4 Isolation of CD34+ Progenitor Cells;248
12.2.5;2.5 Isolation of Basophils from CML Blood;248
12.2.6;2.6 Isolation of Basophils from Normal Blood;249
12.2.7;2.7 Isolation and Purification of Mast Cells from Tissues;250
12.3;3. CULTURE TECHNIQUES ;251
12.3.1;3.1 Culture of Basophils from Their Progenitor Cells;251
12.3.2;3.2 Culture of Mature Isolated Basophils (Normal And CML- Derived);252
12.3.3;3.3 Culture of Mast Cells from Their Progenitors;252
12.3.4;3.4 Culture of Isolated Mature Tissue Mast Cells;253
12.4;4. UTILITY OF SYSTEMS;253
12.4.1;4.1 Basic Science and Cytokine Research;253
12.4.2;4.2 Clinical Immunology and Allergy Research;254
12.4.3;4.3 Pharmacology and Toxicology;254
12.5;5. CONCLUDING REMARKS;255
12.6;LITERATURE CITED;255
13;Purification and Culture of Erythroid Progenitor Cells;260
13.1;1. INTRODUCTION;260
13.2;2. PURIFICATION OF BFU-E;262
13.2.1;2.1 High Purity Method of BFU-E Purification (Method 1) [ 9,10];262
13.2.2;2.2 High Yield Method of BFU-E Purification (Method II) [ 3,14];269
13.2.3;2.3 Purification of BFU-E Using Magnetic Microbeads ( Method 111);270
13.3;3. IN VITRO GENERATION OF CFU-E FROM PURIFIED BFU-E;273
13.3.1;3.1 Methylcellulose Cell Culture for Generation of CFUE [8];273
13.3.2;3.2 CFU- E Generation from BFU- E in Liquid Suspension Culture;274
13.4;4. ERYTHROID PROGENITOR CELL ASSAYS ;275
13.4.1;4.1 Semi-Solid Culture of BFU-E and CFU-E;275
13.4.2;4.2 Determination of Cell Purity in Liquid Culture;277
13.4.3;4.3 Morphology of BFU-E and CFU-E;277
13.5;5. UTILITY OF PURIFICATION PROCEDURES ;278
13.5.1;5.1 Transitional Change of Growth Factor Requirements and Receptors;278
13.5.2;5.2 Studies of Surface Receptors;281
13.5.3;5.3 Studies on Apoptosis;282
13.5.4;5.4 Studies on Signal Transduction and Protein Phosphorylation;283
13.6;6. CONCLUSIONS;285
13.7;ACKNOWLEDGMENTS;285
13.8;REFERENCES;286
14;In Vitro Development of Megakaryocytes and Platelets;288
14.1;1. INTRODUCTION;288
14.2;2. TISSUE PROCUREMENT AND PROCESSING;292
14.2.1;2.1 Isolation of Lineage-Light-Density CB Cells;292
14.2.2;2.2 Isolation of Lineage-Light-Density Fetal Liver Cells;294
14.2.3;2.3 Isolation of Lineage-Light-Density FBM Cells;297
14.2.4;2.4 Preparation of Cells for FACS;298
14.3;3. ASSAY TECHNIQUES ;301
14.3.1;3.1 Flow Cytometric Analysis of Cell Surface Markers;301
14.3.2;3.2 Flow Cytometric Analysis of DNA Content;302
14.4;4. CULTURE TECHNIQUES;304
14.4.1;4.1 Serum- Deprived Media;305
14.4.2;4.2 Colony-Forming Unit Assay;306
14.5;5. UTILITY OF SYSTEMS;308
14.6;6. CONCLUDING REMARKS;310
14.7;LITERATURE CITED;311
15;Perspectives, Ethics and Clinical Issues in the Use of Primary Human Cells;317
15.1;1. INTRODUCTION;317
15.2;2. HISTORICAL AND LEGAL PERSPECTIVES;318
15.2.1;2.1 Whole Organ Transplantation;318
15.2.2;2.2 Tissue, Cell and Gene Transplantation;323
15.3;3. ETHICAL ISSUES AND RESPONSIBILITIES;327
15.3.1;3.1 Acquisition and Distribution;327
15.3.2;3.2 Selection of Donors and Recipients;330
15.3.3;3.3 Genetic Manipulation;333
15.4;4. HARVESTING, PREPARATION AND BANKING ISSUES ;334
15.4.1;4.1 Safety and Efficacy of the Products ;334
15.4.2;4.2 Ownership and Cells, Tissues and Organs as Commodities;336
15.5;5. SUMMARY;337
15.6;REFERENCES;338