E-Book, Englisch, 628 Seiten
Reihe: ISSN
E-Book, Englisch, 628 Seiten
Reihe: ISSN
ISBN: 978-0-08-088525-4
Verlag: Elsevier Reference Monographs
Format: PDF
Kopierschutz: Adobe DRM (»Systemvoraussetzungen)
This volume, as part of the Reliable Lab Solutions series, delivers busy researchers a handy, time-saving source for the best methods and protocols in stem cells.
* Provides powerful research opportunities for those interested in perusing work in pluripotent stem cells, disease modeling, and other aspects of basic stem cell research
* Refines, organizes and updates popular methods from flagship series, Methods in Enzymology
*Highlights top downloads, enhanced with author tips and tricks and pitfalls to avoid
Autoren/Hrsg.
Weitere Infos & Material
1;Front Cover;1
2;Essential Stem Cell Methods;4
3;Copyright Page;5
4;Contents;6
5;Contributors;14
6;Preface;20
7;Section I: Organ-Derived Stem Cells;22
7.1;Chapter 1: Neural Stem Cells: Isolation and Characterization;24
7.1.1;1. Introduction;25
7.1.2;2. Reagents and Instrumentation;26
7.1.3;3. Methods;29
7.1.4;Acknowledgments;42
7.1.5;References;42
7.2;Chapter 2: Neural Stem Cells and Their Manipulation;44
7.2.1;1. Introduction;45
7.2.2;2. Adult Niches for Stem Cells In Vivo;46
7.2.3;3. In Vitro Manipulation of NSCs;48
7.2.4;4. Conclusions and Projections;60
7.2.5;5. Sample Protocols for the Culture and Characterization of NSCs;61
7.2.6;6. Culture of Rodent/Murine NSCs;62
7.2.7;7. Culture of Human NSCs;64
7.2.8;8. ICC of Markers to Identify Stem Cells and Differentiation Products;66
7.2.9;Acknowledgments;66
7.2.10;References;66
7.3;Chapter 3: Retinal Stem Cells;74
7.3.1;1. Introduction;75
7.3.2;2. Materials and Methods;81
7.3.3;3. Overview;89
7.3.4;References;90
7.4;Chapter 4: Dental Pulp Stem Cells;94
7.4.1;1. Introduction;95
7.4.2;2. Identification of Dental Pulp Stem Cells (DPSCs);96
7.4.3;3. Isolation of DPSCs;98
7.4.4;4. Differentiation of DPSCs;100
7.4.5;References;104
7.5;Chapter 5: Mouse Spermatogonial Stem Cells: Culture and Transplantation;108
7.5.1;1. Introduction;109
7.5.2;2. Spermatogonial Stem Cells (SSCs);110
7.5.3;3. SSC Transplantation;112
7.5.4;4. SSC Culture;119
7.5.5;5. siRNA Transfection of Cultured Mouse SSCs;126
7.5.6;6. Implications;130
7.5.7;Acknowledgments;130
7.5.8;References;131
7.6;Chapter 6: Stem Cells in the Adult Lung;134
7.6.1;1. Introduction;135
7.6.2;2. Anatomical and Cellular Diversity of the Adult Lung;136
7.6.3;3. Stem Cell Phenotypes and Niches in the Adult Lung;138
7.6.4;4. In Vivo Injury Models of the Lung;142
7.6.5;5. Ex Vivo Epithelial Tracheal Xenograft Model to Study Stem Cells Expansion in the Proximal Airway;149
7.6.6;6. In Vitro Colony-Forming Efficiency (CFE) Assay to Characterize Stem/progenitor Cell Populations in Conducting Airway Epithel;157
7.6.7;7. Models to Study Stem/Progenitor Cells of Airway SMG;162
7.6.8;Acknowledgments;164
7.6.9;References;164
7.7;Chapter 7: Pancreatic Cells and Their Progenitors;170
7.7.1;1. Introduction;171
7.7.2;2. Pancreas Development;172
7.7.3;3. Origins of Beta Cells During Postnatal Life;174
7.7.4;4. Preexisting Beta Cells;175
7.7.5;5. Ducts;176
7.7.6;6. Acini;176
7.7.7;7. Bone Marrow Cells;177
7.7.8;8. Adult Stem Cells;177
7.7.9;9. Dedifferentiation of Beta Cells;178
7.7.10;10. Summary and Perspective;178
7.7.11;11. Methods: Design of a Lineage Tracing Experiment in Mice;179
7.7.12;References;181
7.8;Chapter 8: Pluripotent Stem Cells from Germ Cells: Derivation and Maintenance;186
7.8.1;1. Introduction;187
7.8.2;2. Germ Cell Development;188
7.8.3;3. EGC Derivation;189
7.8.4;4. Characterization of EG Cultures;198
7.8.5;5. EB Formation and Analysis;203
7.8.6;6. EB-Derived Cell Formation;204
7.8.7;Acknowledgments;206
7.8.8;References;206
7.9;Chapter 9: Pluripotent Stem Cells from Amniotic Fluid and Placenta;212
7.9.1;1. Introduction;213
7.9.2;2. Amniotic Fluid and Placenta in Developmental Biology;213
7.9.3;3. Amniotic and Placental Cells for Therapy-A New Connection;214
7.9.4;4. Isolation and Characterization of Progenitor Cells;215
7.9.5;5. Differentiation of Amniotic and Placental-Derived Progenitor Cells;216
7.9.6;6. Conclusion;220
7.9.7;References;220
7.10;Chapter 10: Hematopoietic Stem and Progenitor Cells from Cord Blood;224
7.10.1;1. Introduction: Cord Blood Transplantation;225
7.10.2;2. Methodologies for Assessing Hematopoietic Progenitor and Stem Cells, as Well as EPC, Present in Human Cord Blood;232
7.10.3;3. Uses of Above Assays;248
7.10.4;References;250
7.11;Chapter 11: Hematopoietic Stem Cells from Bone Marrow: Purification and Functional Analysis;258
7.11.1;1. Introduction;259
7.11.2;2. Other HSC Surface Markers: Tie-2, Endoglin, and the SLAM Family Receptors;260
7.11.3;3. Fluorescent Dye Efflux in HSCs and the SPKLS;260
7.11.4;4. Characteristics of HSCs from Different Purification Schemes;263
7.11.5;5. Protocol of HSC Sorting with Hoechst 33342 Staining (SP Population);264
7.11.6;References;268
7.12;Chapter 12: Microarray Analysis of Stem Cells and Their Differentiation;270
7.12.1;1. Introduction;271
7.12.2;2. Overview of Microarray Technology;271
7.12.3;3. Experimental Design;274
7.12.4;4. Confirmation Studies;291
7.12.5;5. Examples of Microarray Experiments for Stem Cell Biology and Differentiation;292
7.12.6;6. Identification of "Stemness";292
7.12.7;7. Differentiation;293
7.12.8;8. Stem Cell Niches;294
7.12.9;9. Future Directions;296
7.12.10;References;296
7.13;Chapter 13: Tissue Engineering Using Adult Stem Cells;302
7.13.1;1. Introduction;303
7.13.2;2. Biomaterials;305
7.13.3;3. Angiogenic Factors;307
7.13.4;4. Adult Stem Cells for Tissue Engineering;308
7.13.5;5. Conclusion;313
7.13.6;References;313
7.14;Chapter 14: Tissue Engineering Using Mesenchymal Stem Cells;318
7.14.1;1. MSCs: Definition and Therapeutic Promise;319
7.14.2;2. Isolation and Expansion of MSCs;321
7.14.3;3. Multilineage Differentiation of MSCs;324
7.14.4;4. Clinical Translation of MSC-Based Therapies;331
7.14.5;5. Conclusions;333
7.14.6;Acknowledgments;334
7.14.7;References;334
8;Section II: Embryonic Stem Cells and Their Derivatives;340
8.1;Chapter 15: Mouse Embryonic Stem Cells;342
8.1.1;1. Historical Overview;343
8.1.2;2. Factors Affecting the Effi;ciency of Mouse ES Cell Establishment;345
8.1.3;3. Factors Affecting the Contribution of Mouse ES Cell to Chimeric Embryos;346
8.1.4;4. Critical Events During Mouse ES Cell Establishment;347
8.1.5;5. Freezing of ES Cell Lines;351
8.1.6;6. Characterization;351
8.1.7;7. Protocols;353
8.1.8;Acknowledgment;358
8.1.9;References;358
8.2;Chapter 16: Human Embryo Culture;362
8.2.1;1. Introduction;363
8.2.2;2. Human Embryo Development;363
8.2.3;3. Embryo Biopsy;366
8.2.4;4. Human Embryo Culture;368
8.2.5;5. Results;372
8.2.6;References;375
8.3;Chapter 17: Human Embryonic Stem Cells: Derivation and Maintenance;380
8.3.1;1. Introduction;381
8.3.2;2. Derivation of hES Cell Lines;381
8.3.3;3. Maintenance of hES Cells;386
8.3.4;4. Conclusion;389
8.3.5;Acknowledgments;389
8.3.6;References;390
8.4;Chapter 18: Human Embryonic Stem Cells: Characterization and Evaluation;394
8.4.1;1. Introduction;395
8.4.2;2. Characterization of Undifferentiated hESCs;396
8.4.3;3. Conclusion;410
8.4.4;Acknowledgments;410
8.4.5;References;410
8.5;Chapter 19: Human Embryonic Stem Cells: Feeder-Free Culture ;416
8.5.1;1. Introduction;417
8.5.2;2. Methods for Feeder Layer-Free Culture of hESCs;422
8.5.3;Acknowledgments;425
8.5.4;References;425
8.6;Chapter 20: Neural Stem Cells, Neurons, and Glia from Embryonic Stem Cells;428
8.6.1;1. Introduction;429
8.6.2;2. Protocols;430
8.6.3;3. Summary;442
8.6.4;4. Media and Reagents;443
8.6.5;Acknowledgments;444
8.6.6;References;445
8.7;Chapter 21: Hematopoietic Cell Differentiation from Embryonic Stem Cells;446
8.7.1;1. Introduction;447
8.7.2;2. Methods;457
8.7.3;References;472
8.8;Chapter 22: Cardiomyocyte Differentiation from Embryonic Stem Cells;480
8.8.1;1. Introduction;481
8.8.2;2. Embryoid Body Generation;482
8.8.3;3. Cardiomyocyte Differentiation;491
8.8.4;4. Enrichment of Cardiomyocytes;492
8.8.5;5. Conclusion;494
8.8.6;References;495
8.9;Chapter 23: Insulin-Producing Cells from Mouse Embryonic Stem Cells;498
8.9.1;1. Introduction;499
8.9.2;2. Materials and Methods;506
8.9.3;3. Results;514
8.9.4;4. Summary;515
8.9.5;References;517
8.10;Chapter 24: Transgene Expression and RNA Interference in Embryonic Stem Cells;522
8.10.1;1. Retrovirus Expression Vectors and ESCs;523
8.10.2;2. RNA Interference and ESCs;524
8.10.3;3. siRNA Expression Vector Design;525
8.10.4;4. Retrovirus Production;526
8.10.5;5. Retroviral and Lentiviral Gene Transfer into Mouse and Human ESCs;527
8.10.6;6. Transgene and siRNA Expression in Mouse and hESCs;528
8.10.7;7. Biotechnological and Medical Applications;532
8.10.8;Acknowledgments;533
8.10.9;References;533
8.11;Chapter 25: Lentiviral Vector-Mediated Gene Delivery into Human Embryonic Stem Cells;538
8.11.1;1. Introduction;540
8.11.2;2. Design of HIV-1 Based Vectors for Transduction of hESCs;544
8.11.3;3. Generation of Recombinant Viral Particles;548
8.11.4;4. Transduction of hESCs;550
8.11.5;5. Measurement of Transduction Efficiency;553
8.11.6;6. Enrichment for Transduced hESCs Expressing High Levels of the Transgene;554
8.11.7;7. Determination of Viral Titer;556
8.11.8;References;557
8.12;Chapter 26: Engineering Embryonic Stem Cells with Recombinase Systems;562
8.12.1;1. Introduction;563
8.12.2;2. Site-Specific Recombination;564
8.12.3;3. Designing Substrates for Site-Specific Recombination;568
8.12.4;4. Generation of Conditional Alleles;574
8.12.5;5. Recombinase Mediated Cassette Exchange (RMCE);579
8.12.6;6. Molecular Switches;580
8.12.7;7. Protocols;583
8.12.8;Acknowledgments;592
8.12.9;References;593
8.13;Chapter 27: Tissue Engineering Using Embryonic Stem Cells;600
8.13.1;1. Introduction;601
8.13.2;2. Special Considerations when using hESCs as the Cell Source for TE;602
8.13.3;3. Growing hESCs in Defined Animal-Free Conditions;602
8.13.4;4. Obtaining the Desired Cell Population;603
8.13.5;5. Choosing the Right Scaffold;604
8.13.6;6. Scaling-Up a Regulable Bioprocess;604
8.13.7;7. hESC-Derived Connective Tissue Progenitors for TE;605
8.13.8;8. Culture and Maintenance of hESC on MEF Feeders;605
8.13.9;9. Harvesting Samples for Analyses;610
8.13.10;References;614
9;Index;616
