Essential Stem Cell Methods (eBook)

Irina Klimanskaya, Robert Lanza (Herausgeber)

eBook Download: PDF

2011 | 1. Auflage
628 Seiten
Elsevier Science (Verlag)
978-0-08-088525-4 (ISBN)

Infos

This is a fast-moving field, and these detailed methods will help drive advances in stem cell research. The editors have hand selected step-by-step methods from researchers with extensive reputations and expertise.
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

Inhalt

Front Cover 1
Essential Stem Cell Methods 4
Copyright Page 5
Contents 6
Contributors 14
Preface 20
Section I: Organ-Derived Stem Cells 22
Chapter 1: Neural Stem Cells: Isolation and Characterization 24
1. Introduction 25
2. Reagents and Instrumentation 26
3. Methods 29
Acknowledgments 42
References 42
Chapter 2: Neural Stem Cells and Their Manipulation 44
1. Introduction 45
2. Adult Niches for Stem Cells In Vivo 46
3. In Vitro Manipulation of NSCs 48
4. Conclusions and Projections 60
5. Sample Protocols for the Culture and Characterization of NSCs 61
6. Culture of Rodent/Murine NSCs 62
7. Culture of Human NSCs 64
8. ICC of Markers to Identify Stem Cells and Differentiation Products 66
Acknowledgments 66
References 66
Chapter 3: Retinal Stem Cells 74
1. Introduction 75
2. Materials and Methods 81
3. Overview 89
References 90
Chapter 4: Dental Pulp Stem Cells 94
1. Introduction 95
2. Identification of Dental Pulp Stem Cells (DPSCs) 96
3. Isolation of DPSCs 98
4. Differentiation of DPSCs 100
References 104
Chapter 5: Mouse Spermatogonial Stem Cells: Culture and Transplantation 108
1. Introduction 109
2. Spermatogonial Stem Cells (SSCs) 110
3. SSC Transplantation 112
4. SSC Culture 119
5. siRNA Transfection of Cultured Mouse SSCs 126
6. Implications 130
Acknowledgments 130
References 131
Chapter 6: Stem Cells in the Adult Lung 134
1. Introduction 135
2. Anatomical and Cellular Diversity of the Adult Lung 136
3. Stem Cell Phenotypes and Niches in the Adult Lung 138
4. In Vivo Injury Models of the Lung 142
5. Ex Vivo Epithelial Tracheal Xenograft Model to Study Stem Cells Expansion in the Proximal Airway 149
6. In Vitro Colony-Forming Efficiency (CFE) Assay to Characterize Stem/progenitor Cell Populations in Conducting Airway Epithel 157
7. Models to Study Stem/Progenitor Cells of Airway SMG 162
Acknowledgments 164
References 164
Chapter 7: Pancreatic Cells and Their Progenitors 170
1. Introduction 171
2. Pancreas Development 172
3. Origins of Beta Cells During Postnatal Life 174
4. Preexisting Beta Cells 175
5. Ducts 176
6. Acini 176
7. Bone Marrow Cells 177
8. Adult Stem Cells 177
9. Dedifferentiation of Beta Cells 178
10. Summary and Perspective 178
11. Methods: Design of a Lineage Tracing Experiment in Mice 179
References 181
Chapter 8: Pluripotent Stem Cells from Germ Cells: Derivation and Maintenance 186
1. Introduction 187
2. Germ Cell Development 188
3. EGC Derivation 189
4. Characterization of EG Cultures 198
5. EB Formation and Analysis 203
6. EB-Derived Cell Formation 204
Acknowledgments 206
References 206
Chapter 9: Pluripotent Stem Cells from Amniotic Fluid and Placenta 212
1. Introduction 213
2. Amniotic Fluid and Placenta in Developmental Biology 213
3. Amniotic and Placental Cells for Therapy-A New Connection 214
4. Isolation and Characterization of Progenitor Cells 215
5. Differentiation of Amniotic and Placental-Derived Progenitor Cells 216
6. Conclusion 220
References 220
Chapter 10: Hematopoietic Stem and Progenitor Cells from Cord Blood 224
1. Introduction: Cord Blood Transplantation 225
2. Methodologies for Assessing Hematopoietic Progenitor and Stem Cells, as Well as EPC, Present in Human Cord Blood 232
3. Uses of Above Assays 248
References 250
Chapter 11: Hematopoietic Stem Cells from Bone Marrow: Purification and Functional Analysis 258
1. Introduction 259
2. Other HSC Surface Markers: Tie-2, Endoglin, and the SLAM Family Receptors 260
3. Fluorescent Dye Efflux in HSCs and the SPKLS 260
4. Characteristics of HSCs from Different Purification Schemes 263
5. Protocol of HSC Sorting with Hoechst 33342 Staining (SP Population) 264
References 268
Chapter 12: Microarray Analysis of Stem Cells and Their Differentiation 270
1. Introduction 271
2. Overview of Microarray Technology 271
3. Experimental Design 274
4. Confirmation Studies 291
5. Examples of Microarray Experiments for Stem Cell Biology and Differentiation 292
6. Identification of "Stemness" 292
7. Differentiation 293
8. Stem Cell Niches 294
9. Future Directions 296
References 296
Chapter 13: Tissue Engineering Using Adult Stem Cells 302
1. Introduction 303
2. Biomaterials 305
3. Angiogenic Factors 307
4. Adult Stem Cells for Tissue Engineering 308
5. Conclusion 313
References 313
Chapter 14: Tissue Engineering Using Mesenchymal Stem Cells 318
1. MSCs: Definition and Therapeutic Promise 319
2. Isolation and Expansion of MSCs 321
3. Multilineage Differentiation of MSCs 324
4. Clinical Translation of MSC-Based Therapies 331
5. Conclusions 333
Acknowledgments 334
References 334
Section II: Embryonic Stem Cells and Their Derivatives 340
Chapter 15: Mouse Embryonic Stem Cells 342
1. Historical Overview 343
2. Factors Affecting the Effi ciency of Mouse ES Cell Establishment
3. Factors Affecting the Contribution of Mouse ES Cell to Chimeric Embryos 346
4. Critical Events During Mouse ES Cell Establishment 347
5. Freezing of ES Cell Lines 351
6. Characterization 351
7. Protocols 353
Acknowledgment 358
References 358
Chapter 16: Human Embryo Culture 362
1. Introduction 363
2. Human Embryo Development 363
3. Embryo Biopsy 366
4. Human Embryo Culture 368
5. Results 372
References 375
Chapter 17: Human Embryonic Stem Cells: Derivation and Maintenance 380
1. Introduction 381
2. Derivation of hES Cell Lines 381
3. Maintenance of hES Cells 386
4. Conclusion 389
Acknowledgments 389
References 390
Chapter 18: Human Embryonic Stem Cells: Characterization and Evaluation 394
1. Introduction 395
2. Characterization of Undifferentiated hESCs 396
3. Conclusion 410
Acknowledgments 410
References 410
Chapter 19: Human Embryonic Stem Cells: Feeder-Free Culture 416
1. Introduction 417
2. Methods for Feeder Layer-Free Culture of hESCs 422
Acknowledgments 425
References 425
Chapter 20: Neural Stem Cells, Neurons, and Glia from Embryonic Stem Cells 428
1. Introduction 429
2. Protocols 430
3. Summary 442
4. Media and Reagents 443
Acknowledgments 444
References 445
Chapter 21: Hematopoietic Cell Differentiation from Embryonic Stem Cells 446
1. Introduction 447
2. Methods 457
References 472
Chapter 22: Cardiomyocyte Differentiation from Embryonic Stem Cells 480
1. Introduction 481
2. Embryoid Body Generation 482
3. Cardiomyocyte Differentiation 491
4. Enrichment of Cardiomyocytes 492
5. Conclusion 494
References 495
Chapter 23: Insulin-Producing Cells from Mouse Embryonic Stem Cells 498
1. Introduction 499
2. Materials and Methods 506
3. Results 514
4. Summary 515
References 517
Chapter 24: Transgene Expression and RNA Interference in Embryonic Stem Cells 522
1. Retrovirus Expression Vectors and ESCs 523
2. RNA Interference and ESCs 524
3. siRNA Expression Vector Design 525
4. Retrovirus Production 526
5. Retroviral and Lentiviral Gene Transfer into Mouse and Human ESCs 527
6. Transgene and siRNA Expression in Mouse and hESCs 528
7. Biotechnological and Medical Applications 532
Acknowledgments 533
References 533
Chapter 25: Lentiviral Vector-Mediated Gene Delivery into Human Embryonic Stem Cells 538
1. Introduction 540
2. Design of HIV-1 Based Vectors for Transduction of hESCs 544
3. Generation of Recombinant Viral Particles 548
4. Transduction of hESCs 550
5. Measurement of Transduction Efficiency 553
6. Enrichment for Transduced hESCs Expressing High Levels of the Transgene 554
7. Determination of Viral Titer 556
References 557
Chapter 26: Engineering Embryonic Stem Cells with Recombinase Systems 562
1. Introduction 563
2. Site-Specific Recombination 564
3. Designing Substrates for Site-Specific Recombination 568
4. Generation of Conditional Alleles 574
5. Recombinase Mediated Cassette Exchange (RMCE) 579
6. Molecular Switches 580
7. Protocols 583
Acknowledgments 592
References 593
Chapter 27: Tissue Engineering Using Embryonic Stem Cells 600
1. Introduction 601
2. Special Considerations when using hESCs as the Cell Source for TE 602
3. Growing hESCs in Defined Animal-Free Conditions 602
4. Obtaining the Desired Cell Population 603
5. Choosing the Right Scaffold 604
6. Scaling-Up a Regulable Bioprocess 604
7. hESC-Derived Connective Tissue Progenitors for TE 605
8. Culture and Maintenance of hESC on MEF Feeders 605
9. Harvesting Samples for Analyses 610
References 614
Index 616

Produkt-Details

• Erscheint lt. Verlag: 13.10.2011
• Sprache: englisch
• Themenwelt: Sachbuch/Ratgeber
• Themenwelt: Naturwissenschaften ► Biologie ► Zellbiologie
• Themenwelt: Technik
• ISBN-10: 0-08-088525-X / 008088525X
• ISBN-13: 978-0-08-088525-4 / 9780080885254