Koller / Palsson / Masters Primary Mesenchymal Cells

1. Auflage 2006
ISBN: 978-0-306-46870-4
Verlag: Springer Netherlands
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 5, 256 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;Contents;6
2;Introduction;7
3;Foreword to the Series;8
4;Articular Cartilage;9
4.1;1. INTRODUCTION;9
4.2;2. TISSUE PROCUREMENT AND PROCESSING;11
4.3;3. ASSAY TECHNIQUES;12
4.3.1;3.1 Chondrocyte re-differentiation in semi-solid media;13
4.3.2;3.2 Agarose differentiation assay;13
4.3.3;3.3 Alginate differentiation assay;13
4.3.4;3.4 Analysis of gene expression;14
4.3.5;3.5 RNA Analysis;14
4.3.6;3.6 RNase protection assay (RPA);14
4.3.7;3.7 Immunohistochemical Analysis;14
4.4;4. CULTURE TECHNIQUES;15
4.4.1;4.1 Anchorage Dependent Expansion in Monolayer;15
4.4.2;4.2 Autologous human serum (AHS) versus fetal bovine serum (FBS);15
4.4.3;4.3 Impact of Donor Age on Chondrocyte Re-differentiation;16
4.4.4;4.4 Development of a defined medium for articular chondrocyte culture;16
4.4.5;4.5 Re-differentiation of articular chondrocytes cultured in defined medium;19
4.5;5. UTILITY OF THE SYSTEM;19
4.6;6. CONCLUDING REMARKS;21
4.6.1;6.1 The Future Leads to Tissue Engineering;21
4.7;REFERENCES;22
5;Tendon and Ligaments;25
5.1;1. INTRODUCTION;25
5.2;2. TISSUE PROCUREMENT AND PROCESSING;27
5.3;3. ASSAY TECHNIQUES;29
5.4;4. CULTURE TECHNIQUES;30
5.5;5. UTILITY OF THE SYSTEM;31
5.6;6. CONCLUDING REMARKS;32
5.7;REFERENCES;32
6;Periodontal Ligaments;34
6.1;1. INTRODUCTION;34
6.2;2. TISSUE PROCUREMENT;36
6.3;3. ASSAY TECHNIQUES;40
6.4;4. CULTURE TECHNIQUES;41
6.5;5. UTILITY OF THE SYSTEM;45
6.6;6. CONCLUDING REMARKS;46
6.7;REFERENCES;46
7;Vascular Smooth Muscle;49
7.1;1. INTRODUCTION;49
7.1.1;1.1 Location of VSMCs in the blood vessel wall;50
7.1.2;1.2 VSMC phenotype and importance in disease;51
7.1.3;1.3 VSMC heterogeneity and tissue culture models;51
7.1.4;1.4 Importance of using human VSMC cultures;54
7.2;2. TISSUE PROCUREMENT AND PROCESSING;54
7.2.1;2.1 Obtaining human blood vessels;54
7.2.2;2.2 Processing tissue for VSMC isolation;55
7.2.3;2.3 Isolation of VSMCs from the micro-vasculature (pericytes);57
7.2.4;2.4 Isolation of VSMCs from diseased vessels;57
7.3;3. CULTURE TECHNIQUES;58
7.3.1;3.1 Media and supplements;58
7.3.2;3.2 Culture vessels;59
7.3.3;3.3 Passaging of cultures;59
7.4;4. ASSAY TECHNIQUES;60
7.4.1;4.1 Identification of VSMCs;60
7.4.2;4.2 Cell markers;61
7.4.3;4.3 Growth;63
7.5;5. UTILITY OF SYSTEM;63
7.5.1;5.1 VSMC growth and migration;63
7.5.2;5.2 VSMC phenotypic diversity;64
7.5.3;5.3 VSMC contraction/differentiation;65
7.5.4;5.4 Human VSMCs and gene transfer;65
7.6;6. SUMMARY AND CONCLUDING REMARKS;66
7.7;REFERENCES;66
8;Skeletal Muscle;71
8.1;1. INTRODUCTION;71
8.1.1;1.1 Control of Muscle Development in Vertebrates;72
8.1.2;1.2 Regulation of Muscle Cell Determination and Migration;72
8.1.3;1.3 Regeneration of Skeletal Muscle;74
8.2;2. TISSUE PROCUREMENT AND PROCESSING;75
8.2.1;2.1 Tissue Procurement;75
8.2.2;2.2 Tissue Processing;76
8.2.3;2.3 Donor-to-Donor and Muscle-to-Muscle Variability;79
8.3;3. CULTURE TECHNIQUES;80
8.3.1;3.1 Choice of Culture Medium;80
8.3.2;3.2 Coating of Growth Surfaces;81
8.3.3;3.3 Enrichment of the Cell Population for Muscle Cells;83
8.3.4;3.4 Passaging and Conservation;84
8.3.5;3.5 Extending the Life Span of Muscle Cells;85
8.4;4. ASSAY TECHNIQUES;85
8.4.1;4.1 Morphology;86
8.4.2;4.2 Degree of Differentiation;88
8.4.3;4.3 Enhancement of Differentiation by Innervation;91
8.4.4;4.4 Gene Delivery to Cultured Muscle Cells;92
8.4.5;4.5 Making Your Own Muscle Cells;92
8.4.6;4.6 Human Skeletal Muscle Cell Lines;93
8.5;5. CLINICAL USE OF MYOBLASTS;94
8.5.1;5.1 Transplantation of Myoblasts;94
8.5.2;5.2 Myoblasts and Gene Therapy;94
8.5.3;5.3 Myoblasts and Treatment of Heart Failure;95
8.6;6. CONCLUDING REMARKS;96
8.7;REFERENCES;97
9;Cardiomyocytes;108
9.1;1. INTRODUCTION;108
9.1.1;1.1 Cardiomyocyte Development and Differentiation;108
9.1.2;1.2 Cardiomyocyte Cultures;110
9.2;2. TISSUE PROCUREMENT AND PROCESSING;112
9.2.1;2.1 Human Myocardial Tissue;112
9.2.2;2.2 Cell Isolation;112
9.3;3. ASSAY TECHNIQUES;115
9.3.1;3.1 Morphology;115
9.3.2;3.2 Growth Characteristics;116
9.3.3;3.3 Myocardial Contractile Proteins;120
9.3.4;3.4 Cardiac Protein Gene Expression;120
9.3.5;3.5 Cellular Ultrastructure;121
9.3.6;3.6 Biochemical Identification;121
9.4;4. CULTURE TECHNIQUES;121
9.4.1;4.1 Cell Purification;121
9.4.2;4.2 Cell Culture;122
9.5;5. APPLICATIONS OF A CARDIOMYOCYTE CULTURE SYSTEM;124
9.5.1;5.1 Cardiac Development;124
9.5.2;5.2 Cardiac Physiology;124
9.5.3;5.3 Cardiac Pathology and Cellular Defenses;125
9.6;6. CONCLUSIONS;126
9.7;REFERENCES;126
10;Dermal Fibroblasts;130
10.1;1. INTRODUCTION;130
10.1.1;1.1 Brief history of fibroblast culture;130
10.1.2;1.2 Ease and scope of fibroblast culture;131
10.2;2. FIBROBLAST ISOLATION;132
10.2.1;2.1 Fibroblast populations and tissue types;133
10.3;3. CULTURE;133
10.3.1;3.1 Monolayer culture;133
10.3.2;3.2 Fibroblast Culture Lifespan;139
10.3.3;3.3 Three-dimensional culture;145
10.4;4. ANALYTICAL METHODS FOR THE STUDY OF FIBROBLAST CULTURES;150
10.4.1;4.1 Methods for Determining Cell Proliferation;150
10.4.2;4.2 Mathematical description of culture growth;153
10.4.3;4.3 Viability;154
10.4.4;4.4 Extracellular Matrix Synthesis;155
10.5;5. PRACTICAL APPLICATIONS OF DERMAL FIBROBLAST CULTURES;160
10.5.1;5.1 Studies of the properties of primary cultures;160
10.5.2;5.2 Fibroblast interactions with other cell types and feeder cultures;160
10.5.3;5.3 Tissue engineering of connective tissues other than dermis;161
10.5.4;5.4 Therapeutic and testing applications of threedimensional cultures ;161
10.5.5;5.5 Extracellular matrix production;162
10.5.6;5.6 Gene therapy applications;162
10.6;6. CONCLUSION;163
10.7;ACKNOWLEDGEMENTS;163
10.8;REFERENCES;163
11;Adipose Tissue;178
11.1;1. INTRODUCTION;178
11.1.1;1.1 Organization of human adipose tissue;179
11.1.2;1.2 Adipose tissue function;180
11.2;2. TISSUE PROCUREMENT AND PROCESSING;181
11.2.1;2.1 General comments;181
11.2.2;2.2 Isolation of pre-adipocytes and adipocytes;181
11.3;3. CULTURE TECHNIQUES;182
11.3.1;3.1 Adipose tissue explants;182
11.3.2;3.2 Mature adipocytes;182
11.3.3;3.3 Pre-adipocytes;183
11.3.4;3.4 Pre-adipocyte differentiation;183
11.4;4. ASSAY TECHNIQUES;184
11.4.1;4.1 Morphological features of human pre-adipocyte differentiation;185
11.5;5. UTILITY OF THE SYSTEM;189
11.6;6. SUMMARY;189
11.7;REFERENCES;190
12;Mesenchymal Stem Cells;193
12.1;1. INTRODUCTION;193
12.2;2. TISSUE PROCUREMENT AND PROCESSING;195
12.2.1;2.1 Safety from Biohazards - Universal Precautions;195
12.2.2;2.2 Sources of hMSCs;196
12.2.3;2.3 Marrow Processing;196
12.3;3. CULTURE TECHNIQUES;198
12.3.1;3.1 hMSC Expansion in culture;198
12.3.2;3.2 Freezing cultured hMSCs;199
12.4;4. ASSAY TECHNIQUES;200
12.4.1;4.1 Flow Cytometry;200
12.4.2;4.2 Differentiation Assays;202
12.4.3;4.3 Differentiation Assays;206
12.5;5. UTILITY OF THE SYSTEM;206
12.6;6. CONCLUDING REMARKS;207
12.7;REFERENCES;208
13;Peripheral Blood Fibrocytes;212
13.1;1. INTRODUCTION;212
13.2;2. TISSUE PROCUREMENT AND PROCESSING;216
13.3;3. CULTURE TECHNIQUES;217
13.4;4. ASSAY TECHNIQUES;218
13.5;5. UTILITY OF SYSTEM;220
13.6;6. CONCLUDING REMARKS;221
13.7;REFERENCES;221
14;Osteoblasts;223
14.1;1. INTRODUCTION;223
14.1.1;1.1 Rationale for the use of isolated bone cells;223
14.2;2. TISSUE PROCUREMENT AND PROCESSING;227
14.3;3. CULTURE TECHNIQUES;227
14.4;4. ASSAY TECHNIQUES;232
14.4.1;4.1 HOB Characterization;232
14.5;5. UTILITY OF SYSTEM;236
14.6;6. CONCLUDING REMARKS;238
14.7;ACKNOWLEDGMENTS;238
14.8;REFERENCES;238