Essentials of Stem Cell Biology (eBook)

Front Cover 1
Essentials of Stem Cell Biology 4
Copyright Page 5
Contents 6
Contributors 10
Preface 16
Foreword 18
Why Stem Cell Research 20
A New Path: Induced Pluripotent Stem Cells 22
"Stemness": Definitions, Criteria, and Standards 24
Part I: Introduction to Stem Cells 32
Chapter 1. Pluripotential Stem Cells from Vertebrate Embryos: Present Perspective and Future Challenges 34
INTRODUCTION 34
TERMINOLOGY 35
ES-LIKE CELLS IN OTHER SPECIES 36
EMBRYONIC GERM CELLS 37
FUTURE CHALLENGES 38
BIOLOGY OF ES AND ES-LIKE CELLS 38
STEM CELL THERAPY 40
CONCLUSIONS 42
KEY WORDS 42
FURTHER READING 42
Chapter 2. Embryonic Stem Cells in Perspective 44
FURTHER READING 45
Chapter 3. The Development of Epithelial Stem Cell Concepts 48
INTRODUCTION 48
A DEFINITION OF STEM CELLS 49
HIERARCHICALLY ORGANIZED STEM CELL POPULATIONS 50
SKIN STEM CELLS 51
THE INTESTINAL STEM CELL SYSTEM 53
STEM CELL ORGANIZATION ON THE TONGUE 54
GENERALIZED SCHEME 55
SUMMARY 56
KEY WORDS 57
FURTHER READING 58
Chapter 4. Direct Reprogramming of Somatic Cells to a Pluripotent State 60
INTRODUCTION 60
KEY WORDS 62
FURTHER READING 62
Chapter 5. Clinical Translation of Stem Cells 64
INTRODUCTION 64
CELL PROCESSING AND MANUFACTURING 64
STUDIES WITH STEM CELLS IN ANIMAL MODELS 65
CLINICAL TRIALS WITH STEM CELLS IN PATIENTS 65
CONCLUSIONS 66
FURTHER READING 66
Part II: Basic Biology/Mechanisms 68
Chapter 6. Molecular Basis of Pluripotency 70
INTRODUCTION 70
CELLULAR MODELS OF PLURIPOTENCY 70
THE EMBRYONIC STEM CELL ENVIRONMENT: CYTOKINES AND PLURIPOTENCY 71
CYTOKINE-RECEPTOR BINDING ON ES CELLS: MULTIPLE RELAY STATIONS 72
SIGNAL TRANSDUCTION: CASCADES TO THE STEM CELL NUCLEUS 73
STATS: LATENT TRANSCRIPTION FACTORS TRANSMITTING SIGNALS 73
STAT3 74
SHP-2/ERK SIGNALING 75
FGF4 AND ERK 76
WNT SIGNALING 76
THE TGF& #946
THE PI3K/AKT SIGNALING PATHWAY PARTICIPATES IN ES CELL PROLIFERATION, SURVIVAL, AND PLURIPOTENCY 77
THE NEW PARADIGM OF EMBRYONIC STEM CELL RENEWAL 78
INTRINSIC DETERMINANTS OF PLURIPOTENCY 79
OCT4 IS A KEY TRANSCRIPTION FACTOR FOR PLURIPOTENCY 79
OCT4 EXPRESSION 79
OCT4 ACTIVITY IN ASSAYS 80
REGULATION OF OCT4 EXPRESSION 81
OCT4 TARGET GENES 82
OCT4 DOES NOT PLAY SOLO: SOX2 82
FOXD3 84
NANOG 84
THE CORE TRANSCRIPTIONAL REGULATORY CIRCUITRY CONTROLLING ES CELL IDENTITY 86
THE ROLE OF MICRO-RNAS IN PLURIPOTENCY 86
DGCR8 87
THE CHROMATIN STATUS DEFINING STEM CELL IDENTITY 87
REPROGRAMMING OF SOMATIC CELLS TO A PLURIPOTENT STATE: IPS VERSUS ES 88
CONCLUSIONS 89
KEY TERMS 90
FURTHER READING 90
Chapter 7. Stem Cell Niches 92
INTRODUCTION 92
STEM CELL NICHE HYPOTHESIS 92
STEM CELL NICHES IN THE DROSOPHILA GERMLINE 92
STEM CELL NICHES WITHIN MAMMALIAN TISSUES 96
SUMMARY 101
ACKNOWLEDGMENTS 102
KEY WORDS 103
FURTHER READING 103
Chapter 8. Mechanisms of Stem Cell Self-renewal 104
INTRODUCTION 104
SELF-RENEWAL OF PLURIPOTENT STEM CELLS 104
MOLECULAR MECHANISM FOR RETENTION OF EMBRYONIC STEM CELL SELF-RENEWAL 105
EPIGENETIC REGULATION OF SELF-RENEWAL 109
SELF-RENEWAL OF HUMAN EMBRYONIC STEM CELLS 110
SUMMARY 110
KEY WORDS 110
FURTHER READING 110
Chapter 9. Cell Cycle Regulators in Adult Stem Cells 112
INTRODUCTION 112
CELL CYCLE KINETICS OF STEM CELLS IN VIVO 112
STEM CELL EXPANSION EX VIVO 113
MAMMALIAN CELL CYCLE REGULATION AND CYCLIN-DEPENDENT KINASE INHIBITORS 113
ROLES OF CYCLIN-DEPENDENT KINASE INHIBITORS IN STEM CELL REGULATION 114
SUMMARY 117
ACKNOWLEDGMENTS 117
KEY WORDS 117
FURTHER READING 117
Chapter 10. Epigenetic Mechanisms of Cellular Memory during Development 120
INTRODUCTION 120
CHROMATIN-MODIFYING FACTORS 123
ATPASE CHROMATIN-REMODELING COMPLEXES 125
EPIGENETIC PROCESSES 126
X-CHROMOSOME INACTIVATION 128
MONOALLELIC EXPRESSION: GENOMIC IMPRINTING AND ALLELIC EXCLUSION 131
SUMMARY AND FUTURE DIRECTIONS 133
KEY WORDS 134
FURTHER READING 134
Chapter 11. Cell Fusion and the Differentiated State 136
INTRODUCTION 136
HYBRID CELLS AND DIFFERENTIATED PHENOTYPES 136
HYBRIDS OF PLURIPOTENT CELLS 137
REPROGRAMMING SOMATIC CELL NUCLEI WITH EC, ES, OR EG CELL CYTOPLASM 138
CELL FUSION AND THE DEMONSTRATION OF STEM CELL PLASTICITY 139
SUMMARY 140
ACKNOWLEDGMENTS 140
KEY WORDS 140
FURTHER READING 141
Chapter 12. How Cells Change Their Phenotype 142
INTRODUCTION 142
METAPLASIA AND TRANSDIFFERENTIATION 142
WHY STUDY TRANSDIFFERENTIATION? 142
EXAMPLES OF THE PHENOMENON 142
DEDIFFERENTIATION AS A PREREQUISITE FOR TRANSDIFFERENTIATION 145
HOW TO CHANGE A CELL'S PHENOTYPE EXPERIMENTALLY 146
SUMMARY 147
ACKNOWLEDGMENTS 147
KEY WORDS 147
FURTHER READING 147
Part III: Tissue and Organ Development 148
Chapter 13. Differentiation in Early Development 150
INTRODUCTION 150
PRE-IMPLANTATION DEVELOPMENT 150
FROM IMPLANTATION TO GASTRULATION 156
ACKNOWLEDGMENTS 160
KEY WORDS 160
FURTHER READING 160
Chapter 14. Primordial Germ Cells in Mouse and Human 162
INTRODUCTION 162
ORIGIN OF THE GERM CELL LINEAGE 162
MIGRATION 164
GERM CELLS IN THE GENITAL RIDGE 164
EMBRYONIC GERM CELLS 166
SUMMARY 166
KEY WORDS 167
FURTHER READING 167
Chapter 15. Stem Cells in Extraembryonic Lineages 168
INTRODUCTION 168
TROPHOBLAST LINEAGE 168
TROPHOBLAST STEM CELL LINES 172
EXTRAEMBRYONIC ENDODERM LINEAGE 174
SUMMARY 175
KEY WORDS 175
FURTHER READING 175
Chapter 16. Amniotic Fluid-Derived Pluripotential Cells 176
INTRODUCTION 176
AMNIOTIC FLUID AND AMNIOCENTESIS 176
ISOLATION AND CHARACTERIZATION OF PROGENITOR CELLS 177
DIFFERENTIATION POTENTIAL OF AMNIOTIC PROGENITOR CELLS 177
IN VIVO BEHAVIOR OF AMNIOTIC FLUID STEM CELLS 180
FUTURE DIRECTIONS 180
SUMMARY 180
KEY WORDS 181
FURTHER READING 181
Chapter 17. Cord Blood Hematopoietic Stem and Progenitor Cells 182
INTRODUCTION 182
CHARACTERISTICS AND CRYOPRESERVATION OF CORD BLOOD STEM AND PROGENITOR CELLS 182
CORD BLOOD TRANSPLANTATION PROBLEMS AND POSSIBLE COUNTERMEASURES 183
HOMING OF STEM AND PROGENITOR CELLS 185
SUMMARY 186
KEY WORDS 186
FURTHER READING 186
Chapter 18. Neurogenesis in the Vertebrate Embryo 188
INTRODUCTION 188
EMBRYONIC INDUCTION AND THE ESTABLISHMENT OF NEURAL TISSUE 188
MOLECULAR BASES OF NEURAL INDUCTION 188
NEURAL PATTERNING 191
PRONEURAL GENE EXPRESSION 195
NEGATIVE REGULATORS OF PRONEURAL ACTIVITY 195
REGULATION OF NEURONAL DIFFERENTIATION BY REST/NRSF 197
SWITCH FROM NEUROGENESIS TO GLIOGENESIS 197
SUMMARY 197
KEY WORDS 198
FURTHER READING 198
Chapter 19. The Nervous System 200
INTRODUCTION 200
NEURAL DEVELOPMENT 200
NEURAL STEM CELLS 201
NEURAL DIFFERENTIATION OF MOUSE EMBRYONIC STEM CELLS 201
NEURAL DIFFERENTIATION OF HUMAN AND NONHUMAN PRIMATE ES CELLS 206
DEVELOPMENTAL PERSPECTIVE 207
THERAPEUTIC PERSPECTIVES 207
CONCLUSION 208
KEY WORDS 209
FURTHER READING 209
Chapter 20. Neuronal Progenitors in the Adult Brain: From Development to Regulation 210
INTRODUCTION 210
HISTORY OF STEM CELLS IN THE ADULT CENTRAL NERVOUS SYSTEM 210
GLIAL CHARACTERISTICS OF NEURAL STEM CELLS 210
ADULT NEUROGENESIS IN VIVO 211
SUMMARY 213
KEY WORDS 213
FURTHER READING 213
Chapter 21. Sensory Epithelium of the Eye and Ear: Update of Retinal Stem Cell Research 2003 to the Present 216
INTRODUCTION 216
MUELLER GLIA AS A SOURCE OF NEW RETINAL NEURONS 216
EMBRYONIC STEM CELLS AS A SOURCE OF RETINAL NEURONS 217
KEY WORDS 218
FURTHER READING 219
Chapter 22. Epithelial Hair Follicle Stem Cells 220
INTRODUCTION 220
A BRIEF INTRODUCTION TO MOUSE SKIN ORGANIZATION 220
THE BULGE AS A RESIDENCE OF HAIR FOLLICLE STEM CELLS 222
MODELS OF HAIR FOLLICLE STEM CELL ACTIVATION 224
MOLECULAR FINGERPRINT OF THE BULGE: PUTATIVE STEM CELL MARKERS 225
CELL SIGNALING IN HAIR FOLLICLE STEM CELLS 226
COMMENTARY AND FUTURE DIRECTIONS 227
KEY WORDS 228
FURTHER READING 228
Chapter 23. The Ontogeny of the Hematopoietic System 230
HISTORIC PERSPECTIVE 230
SITES OF INITIATION OF PRIMITIVE AND DEFINITIVE HEMATOPOIESIS AND VASCULOGENESIS 230
STEM CELL MIGRATION TO LATER SITES OF HEMATOPOIESIS 236
CELL MIGRATION TO PRIMARY LYMPHOID ORGANS 237
KEY WORDS 240
FURTHER READING 240
Chapter 24. Hematopoietic Stem Cells 242
EMBRYONIC STEM CELLS AND EMBRYONIC HEMATOPOIESIS 242
BLOOD FORMATION IN EMBRYOID BODIES 243
TRANSFORMATION OF AN EB-DERIVED HSC BY BCR/ABL 243
PROMOTING HEMATOPOIETIC ENGRAFTMENT WITH STAT5 AND HOXB4 243
PROMOTING BLOOD FORMATION IN VITRO WITH EMBRYONIC MORPHOGENS 245
KEY WORDS 245
FURTHER READING 246
Chapter 25. Red Blood Cells 248
ERYTHROPOIESIS 248
ERYTHROCYTES GENERATED FROM ADULT STEM CELLS IN VITRO 248
ERYTHROCYTES GENERATED FROM MOUSE EMBRYONIC STEM CELLS 249
ERYTHROCYTES GENERATED FROM HUMAN EMBRYONIC STEM CELLS 249
ENUCLEATION OF ERYTHROID CELLS 252
SUMMARY 253
KEY WORDS 253
FURTHER READING 253
Chapter 26. Cell Differentiation in the Skeleton 254
INTRODUCTION 254
SKELETOGENESIS 254
CHONDROCYTE DIFFERENTIATION 255
OSTEOBLAST DIFFERENTIATION 256
KEY WORDS 257
FURTHER READING 257
Chapter 27. Human Vascular Progenitor Cells 258
HUMAN VASCULAR DEVELOPMENT, MAINTENANCE AND RENEWAL 258
HUMAN EMBRYONIC STEM CELLS AS A SOURCE FOR VASCULAR PROGENITORS 260
ACKNOWLEDGMENTS 263
KEY WORDS 263
FURTHER READING 263
Chapter 28. Multipotent Adult Progenitor Cells 264
INTRODUCTION 264
ISOLATION AND PHENOTYPIC CHARACTERIZATION OF MULTIPOTENT ADULT PROGENITOR CELLS 264
IN VITRO DIFFERENTIATION POTENTIAL OF MULTIPOTENT ADULT PROGENITOR CELLS 265
IN VIVO ENGRAFTMENT OF MULTIPOTENT ADULT PROGENITOR CELLS 266
MULTIPOTENT ADULT PROGENITOR CELLS IN ISCHEMIA MODELS 266
SYSTEMIC TRANSPLANTATION OF MULTIPOTENT ADULT PROGENITOR CELLS 267
OTHER ADULT STEM CELLS WITH GREATER DIFFERENTIATION POTENTIAL 268
POSSIBLE MECHANISMS UNDERLYING THE GREATER POTENCY OF MULTIPOTENT ADULT PROGENITOR CELLS 268
KEY WORDS 270
FURTHER READING 271
Chapter 29. Mesenchymal Stem Cells 274
INTRODUCTION 274
TISSUE-ENGINEERED REGENERATION 276
ISOLATING AND CULTURING MESENCHYMAL STEM CELLS 277
TISSUE-ENGINEERING USES OF MESENCHYMAL STEM CELLS 277
SUMMARY 279
ACKNOWLEDGMENTS 279
KEY WORDS 279
FURTHER READING 279
Chapter 30. Skeletal Muscle Stem Cells 280
INTRODUCTION 280
SATELLITE CELLS IDENTIFICATION 280
SATELLITE CELL HETEROGENEITY 282
SATELLITE CELL SELF-RENEWAL 283
INTRINSIC REGULATION OF SATELLITE CELLS 284
EXTRINSIC REGULATION OF SATELLITE CELLS 285
SUMMARY 288
ACKNOWLEDGMENTS 288
KEY WORDS 288
FURTHER READING 288
Chapter 31. Stem Cells and the Regenerating Heart 290
INTRODUCTION 290
EVOLVING CONCEPTS OF REGENERATION 292
ACKNOWLEDGMENTS 294
KEY WORDS 294
FURTHER READING 294
Chapter 32. Potential of Embryonic and Induced Pluripotent Stem Cell Differentiation Culture for Vascular Biology 296
INTRODUCTION 296
CULTURES FOR EMBRYONIC STEM CELL DIFFERENTIATION 296
MARKERS FOR DEFINING INTERMEDIATE STAGES DURING ENDOTHELIAL CELL DIFFERENTIATION 297
UTILITY OF EMBRYONIC STEM CELL CULTURE FOR CELL BIOLOGY OF ENDOTHELIAL CELLS 298
ENDOTHELIAL CELL DIVERSIFICATION 299
CONCLUDING REMARKS 301
KEY WORDS 302
FURTHER READING 302
Chapter 33. Cell Lineages and Stem Cells in the Embryonic Kidney 304
INTRODUCTION 304
THE ANATOMY OF KIDNEY DEVELOPMENT 304
GENES THAT CONTROL EARLY KIDNEY DEVELOPMENT 307
THE ESTABLISHMENT OF ADDITIONAL CELL LINEAGES 310
WHAT CONSTITUTES A RENAL STEM CELL? 312
ACKNOWLEDGMENTS 313
KEY WORDS 313
FURTHER READING 313
Chapter 34. Adult Liver Stem Cells 316
INTRODUCTION 316
ORGANIZATION AND FUNCTIONS OF ADULT MAMMALIAN LIVER 316
LIVER STEM CELLS 317
IMMORTAL LIVER PROGENITOR CELL LINES 323
KEY WORDS 329
FURTHER READING 329
Chapter 35. Pancreatic Stem Cells 330
INTRODUCTION 330
DEFINITION OF STEM CELLS AND OF PROGENITOR CELLS 330
PROGENITOR CELLS DURING EMBRYONIC DEVELOPMENT OF THE PANCREAS 331
PROGENITOR CELLS IN THE ADULT PANCREAS 332
FORCING OTHER TISSUES TO ADOPT A PANCREATIC PHENOTYPE 334
IN VITRO STUDIES 335
SUMMARY 336
KEY WORDS 336
FURTHER READING 336
Chapter 36. Stem Cells in the Gastrointestinal Tract 338
INTRODUCTION 338
GASTROINTESTINAL MUCOSA CONTAINS MULTIPLE LINEAGES 339
EPITHELIAL CELL LINEAGES ORIGINATE FROM A COMMON PRECURSOR CELL 339
SINGLE INTESTINAL STEM CELLS REGENERATE WHOLE CRYPTS CONTAINING ALL EPITHELIAL LINEAGES 340
MOUSE AGGREGATION CHIMERAS SHOW THAT INTESTINAL CRYPTS ARE CLONAL POPULATIONS 340
SOMATIC MUTATIONS IN STEM CELLS REVEAL STEM CELL HIERARCHY AND CLONAL SUCCESSION 341
HUMAN INTESTINAL CRYPTS CONTAIN MULTIPLE EPITHELIAL CELL LINEAGES DERIVED FROM A SINGLE STEM CELL 344
BONE MARROW STEM CELLS CONTRIBUTE TO GUT REPOPULATION AFTER DAMAGE 345
GASTROINTESTINAL STEM CELLS OCCUPY A NICHE MAINTAINED BY INTESTINAL SUBEPITHELIAL MYOFIBROBLASTS IN THE LAMINA PROPRIA 348
MANY SIGNALING PATHWAYS REGULATE GASTROINTESTINAL DEVELOPMENT, PROLIFERATION, AND DIFFERENTIATION 350
WNT/& #946
WNT TARGETS EPHB DETERMINE CELL POSITIONING ALONG THE CRYPT AXIS 351
NOTCH SIGNALING IS ALSO TO MAINTAIN THE PROLIFERATIVE COMPARTMENT OF INTESTINAL CRYPTS, AND DETERMINES CELL FATE 352
BONE MORPHOGENIC PATHWAY SIGNALING REGULATES PROLIFERATION ALONG THE INTESTINAL CRYPT AXIS 353
HEDGEHOG SIGNALING PATTERNS THE CRYPT AXIS 353
HOX GENES DEFINE REGIONAL GUT SPECIFICATION 353
GASTROINTESTINAL NEOPLASMS ORIGINATE IN STEM CELL POPULATIONS 353
SUMMARY 357
KEY WORDS 357
FURTHER READING 358
Part IV: Methods 360
Chapter 37. Induced Pluripotent Stem Cell Derivation 362
INTRODUCTION 362
MATERIALS 362
PREPARING TRANSGENE-EXPRESSING LENTIVIRUS 363
REPROGRAMMING HUMAN FORESKIN FIBROBLASTS 366
ACKNOWLEDGMENTS 368
KEY WORDS 368
FURTHER READING 368
Chapter 38. Characteristics and Characterization of Human Pluripotent Stem Cells 370
INTRODUCTION 370
CHARACTERIZATION OF PLURIPOTENT CELLS 372
SUMMARY 373
KEY WORDS 374
FURTHER READING 374
Chapter 39. Isolation and Maintenance of Murine Embryonic Stem Cells 376
INTRODUCTION 376
MAINTENANCE OF EMBRYONIC STEM CELLS 376
MEDIA 377
SERA 377
COLONY-FORMING ASSAY FOR TESTING CULTURE CONDITIONS 377
EMBRYONIC STEM CELL PASSAGE CULTURE 378
ISOLATION OF NEW EMBRYONIC STEM CELL LINES 378
METHOD FOR DERIVATION OF EMBRYONIC STEM CELLS 379
SUMMARY 380
FURTHER READING 380
Chapter 40. Isolation, Characterization and Maintenance of Primate Embryonic Stem Cells 382
INTRODUCTION 382
WHAT ARE PRIMATE EMBRYONIC STEM CELLS? 383
TECHNIQUES FOR THE DERIVATION OF HUMAN EMBRYONIC STEM CELL LINES 384
DERIVATION OF HUMAN EMBRYONIC STEM CELL SUBCLONES 388
METHODS FOR HUMAN EMBRYONIC STEM CELL CULTURE 389
PRIMATE VERSUS MOUSE EMBRYONIC STEM CELLS 391
PRIMATE EMBRYONIC STEM CELL DIFFERENTIATION SYSTEMS 393
ACKNOWLEDGMENTS 393
FURTHER READING 393
Chapter 41. Approaches for Derivation and Maintenance of Human Embryonic Stem Cells: Detailed Procedures and Alternatives 396
INTRODUCTION 396
SETTING UP THE LABORATORY 396
PREPARING AND SCREENING REAGENTS 397
PREPARING PRIMARY MOUSE EMBRYO FIBROBLAST (PMEF) FEEDERS 400
MECHANICAL PASSAGING OF HUMAN EMBRYONIC STEM CELL COLONIES 400
DERIVATION OF HUMAN EMBRYONIC STEM CELLS 402
MAINTENANCE OF ESTABLISHED HUMAN EMBRYONIC STEM CELL CULTURES 403
GLOSSARY TERMS 409
FURTHER READING 409
Chapter 42. Derivation and Differentiation of Human Embryonic Germ Cells 412
INTRODUCTION 412
HUMAN EMBRYONIC GERM CELL DERIVATION 413
EMBRYOID BODY-DERIVED CELLS 418
SUMMARY 421
KEY WORDS 421
FURTHER READING 421
Chapter 43. Growth Factors and the Serum-free Culture of Human Pluripotent Stem Cells 422
INTRODUCTION 422
MOUSE EMBRYONIC STEM CELLS 422
HUMAN EMBRYONAL CARCINOMA CELLS 423
HUMAN EMBRYONIC STEM CELLS 423
EXTRACELLULAR MATRIX COMPONENTS 425
FUTURE PROSPECTS 425
KEY WORDS 426
FURTHER READING 426
Chapter 44. Feeder-free Culture 428
INTRODUCTION 428
FEEDER-FREE CULTURE USING CONDITIONED MEDIUM AND MATRIGEL 428
FEEDER-FREE CULTURE USING CONDITIONED MEDIUM AND MATRIGEL OR LAMININ 431
FEEDER-FREE CULTURE USING DEFINED MEDIA AND MATRIGEL 432
MATERIALS FOR FEEDER-FREE HUMAN EMBRYONIC STEM CELL CULTURE 436
KEY WORDS 438
FURTHER READING 438
Chapter 45. Genetic Manipulation of Human Embryonic Stem Cells 440
INTRODUCTION 440
METHODS OF GENETIC MANIPULATION 440
GENETIC MODIFICATION APPROACHES 441
CONCLUSIONS 446
KEY WORDS 446
FURTHER READING 446
Chapter 46. Homologous Recombination in Human Embryonic Stem Cells 448
INTRODUCTION 448
TARGETED ABLATION OF THE HPRT1 GENE AS A TOOL TO OPTIMIZE HOMOLOGOUS RECOMBINATION EFFICIENCY 448
OCT4 EGFP/NEO KNOCKIN 450
TH EGFP KNOCKIN 451
CONCLUSIONS 452
ACKNOWLEDGMENTS 452
KEY WORDS 452
FURTHER READING 453
Chapter 47. Surface Antigen Markers 454
INTRODUCTION 454
CELL-SURFACE EMBRYONIC ANTIGENS OF THE LABORATORY MOUSE 454
HUMAN EMBRYONAL CARCINOMA AND EMBRYONIC STEM CELLS 455
SUMMARY 456
APPENDIX: METHODS 457
NOTES 458
SOLUTIONS AND NOTES 458
ACKNOWLEDGMENTS 459
KEY WORDS 459
FURTHER READING 459
Chapter 48. Lineage Marking 460
DEFINITIONS 460
QUESTIONS TO ASK 460
MARKERS AND LINEAGE MARKING 461
FUTURE DIRECTIONS 466
ACKNOWLEDGMENTS 466
KEY WORDS 466
FURTHER READING 466
Chapter 49. Genomic Reprogramming 468
INTRODUCTION 468
GENOMIC REPROGRAMMING IN GERM CELLS 468
REPROGRAMMING SOMATIC NUCLEI 472
CONCLUSIONS 472
KEY WORDS 473
FURTHER READING 473
Chapter 50. Isolation and Characterization of Hematopoietic Stem Cells 474
INTRODUCTION 474
ISOLATION OF HEMATOPOIETIC STEM CELLS FROM MICE 474
ISOLATION OF HEMATOPOIETIC STEM CELLS FROM HUMANS 477
HEMATOPOIETIC STEM CELLS AS A PARADIGM FOR STEM CELL BIOLOGY 478
ACKNOWLEDGMENT 478
KEY WORDS 478
FURTHER READING 478
Chapter 51. Microarray Analysis of Stem Cells and Differentiation 480
INTRODUCTION 480
KEY WORDS 487
FURTHER READING 488
Chapter 52. Zebrafish and Stem Cell Research 490
INTRODUCTION 490
THE ZEBRAFISH SYSTEM 490
GENETIC SCREENS IN ZEBRAFISH 492
ZEBRAFISH BLOOD MUTANTS 493
CELL SORTING AND TRANSPLANTATION 494
DISEASE MODELS IN ZEBRAFISH 495
THE FUTURE OF THE SYSTEM 495
ACKNOWLEDGMENTS 496
KEY WORDS 496
FURTHER READING 496
Part V: Applications 498
Chapter 53. Cancer Stem Cells 500
INTRODUCTION 500
CLONALITY AND HETEROGENEITY IN TUMORS 500
THE CANCER STEM CELL MODEL 501
DEFINING FEATURES OF STEM CELLS IN NORMAL TISSUES 502
NOT ALL CANCER CELLS ARE TUMORIGENIC: CLINICAL AND EXPERIMENTAL EVIDENCE 503
SOME CANCERS CAN DIFFERENTIATE INTO NONMALIGNANT CELLS 504
EVIDENCE SUPPORTING THE CANCER STEM CELL MODEL: PROSPECTIVE ISOLATION OF CANCER STEM CELLS 504
ORIGINS OF CANCER STEM CELLS: THE PLASTICITY OF SELF-RENEWAL 507
SIGNALING PATHWAYS IN CANCER STEM CELLS 508
CLINICAL IMPLICATIONS OF CANCER STEM CELLS 509
FUTURE DIRECTIONS 510
FURTHER READING 510
Chapter 54. Neural Stem Cells for Central Nervous System Repair 516
INTRODUCTION 516
THERAPEUTIC POTENTIAL OF NEURAL STEM CELLS 516
GENE THERAPY USING NEURAL STEM CELLS 520
CELL REPLACEMENT USING NEURAL STEM CELLS 521
"GLOBAL" CELL REPLACEMENT USING NEURAL STEM CELLS 521
NEURAL STEM CELLS DISPLAY AN INHERENT MECHANISM FOR RESCUING DYSFUNCTIONAL NEURONS 523
NEURAL STEM CELLS AS THE GLUE THAT HOLDS MULTIPLE THERAPIES TOGETHER 525
SUMMARY 525
KEY WORDS 527
FURTHER READING 527
Chapter 55. Spinal Cord Injury 528
INTRODUCTION 528
PROBLEM 528
SPINAL CORD ORGANIZATION 528
INJURY 528
SPONTANEOUS REGENERATION 529
LIMITATIONS AND APPROACHES TO REPAIR AND REDEFINING GOALS 529
SPINAL CORD DEVELOPMENT 530
EMBRYONIC STEM CELLS 531
EMBRYONIC STEM CELLS AND THE NEURAL LINEAGE 531
EMBRONIC STEM CELL TRANSPLANTATION 531
NOVEL APPROACHES TO CENTRAL NERVOUS SYSTEM REPAIR 536
TOWARD HUMAN TRIALS 537
SUMMARY 537
KEY WORDS 537
FURTHER READING 537
Chapter 56. Use of Embryonic Stem Cells to Treat Heart Disease 538
INTRODUCTION 538
CARDIOMYOCYTE TRANSPLANTATION AS A PARADIGM FOR THE TREATMENT OF DISEASED HEARTS 538
EMBRYONIC STEM CELL-DERIVED CARDIOMYOCYTES, AND FACTORS WHICH ENHANCE CARDIOMYOGENIC DIFFERENTIATION 539
TRANSPLANTATION OF EMBRYONIC STEM CELL-DERIVED CARDIOMYOCYTES IN PRECLINICAL STUDIES, AND CHALLENGES FOR THEIR CLINICAL IMPLEMENTATION 540
SUMMARY 543
ACKNOWLEDGMENTS 543
KEY WORDS 543
FURTHER READING 543
Chapter 57. Insulin-producing Cells Derived from Stem Cells: A Potential Treatment for Diabetes 544
THE NEED FOR INSULIN-PRODUCING CELLS 544
DEFINING & #946
THE POTENTIAL OF EMBRYONIC STEM CELLS AS A SOURCE OF INSULIN-PRODUCING CELLS 545
THE POTENTIAL OF ADULT STEM/PROGENITOR CELLS AS A SOURCE OF INSULIN-PRODUCING CELLS 547
TRANSDIFFERENTIATION OF NON-ISLET CELLS TO ISLET CELLS 550
CONCLUSION ON ADULT SOURCES OF NEW ISLET CELLS 551
ACKNOWLEDGMENTS 551
KEY WORDS 551
FURTHER READING 552
Chapter 58. Burns and Skin Ulcers 554
INTRODUCTION 554
BURNS AND SKIN ULCERS: THE PROBLEM 554
EPIDERMAL STEM CELLS 554
STEM CELLS IN BURNS AND SKIN ULCERS: CURRENT USE 555
FUTURE DEVELOPMENTS 557
SUMMARY 558
ACKNOWLEDGMENTS 559
KEY WORDS 559
FURTHER READING 559
Chapter 59. Stem Cells and Heart Disease 560
THE HEART IS A SELF-RENEWING ORGAN 560
THE HUMAN HEART CONTAINS A POOL OF RESIDENT CARDIAC PROGENITOR CELLS 561
ANATOMICAL DISTRIBUTION OF CARDIAC PROGENITOR CELLS 565
REPAIR OF MYOCARDIAL DAMAGE BY NON-RESIDENT PROGENITOR CELLS 566
REPAIR OF MYOCARDIAL DAMAGE BY RESIDENT PROGENITOR CELLS 568
MYOCARDIAL REGENERATION IN HUMANS 572
KEY WORDS 573
FURTHER READING 573
Chapter 60. Stem Cells for the Treatment of Muscular Dystropy: Therapeutic Perspectives 574
INTRODUCTION 574
MYOBLAST TRANSPLANTATION: REASONS FOR FAILURE AND NEW PERSPECTIVES 575
MYOGENIC STEM CELLS IN THE BONE MARROW AND BONE MARROW TRANSPLANTATION 575
CELLULAR ENVIRONMENT OF A DYSTROPHIC MUSCLE: RESIDENT AND MIGRATORY CELLS 578
FUTURE PERSPECTIVES 579
ACKNOWLEDGMENTS 579
KEY WORDS 579
FURTHER READING 580
Chapter 61. Regeneration of Epidermis from Adult Keratinocyte Stem Cells 582
INTRODUCTION 582
FUTURE DEVELOPMENT 590
SUMMARY 590
ACKNOWLEDGMENTS 590
KEY WORDS 590
FURTHER READING 591
Chapter 62. Orthopedic Applications of Stem Cells 592
INTRODUCTION 592
BONE 593
CARTILAGE 594
MENISCUS 596
LIGAMENTS AND TENDONS 597
SPINE 599
SUMMARY 600
KEY WORDS 600
FURTHER READING 600
Chapter 63. Embryonic Stem Cells in Tissue Engineering 602
INTRODUCTION 602
TISSUE ENGINEERING PRINCIPLES AND PERSPECTIVES 602
LIMITATIONS AND HURDLES OF USING EMBRYONIC STEM CELLS IN TISSUE ENGINEERING 606
CONCLUSIONS AND FUTURE PERSPECTIVES 608
KEY WORDS 608
FURTHER READING 608
Chapter 64. Postnatal Stem Cells in Tissue Engineering 614
INTRODUCTION 614
THE RESERVOIRS OF POSTNATAL STEM CELLS 614
CURRENT APPROACHES TO TISSUE ENGINEERING 616
CONCLUSIONS 620
ACKNOWLEDGMENTS 621
KEY WORDS 621
FURTHER READING 621
Chapter 65. Stem Cell Gene Therapy 622
INTRODUCTION 622
GENE ADDITION 623
GENOME EDITING 624
REQUIREMENTS FOR SUCCESSFUL STEM CELL GENE THERAPY 624
CONCLUSION 627
KEY WORDS 628
FURTHER READING 628
Part VI: Regulation and Ethics 630
Chapter 66. Ethical Considerations 632
INTRODUCTION 632
IS IT MORALLY PERMISSIBLE TO DESTROY A HUMAN EMBRYO? 632
SHOULD WE POSTPONE HUMAN EMBRYONIC STEM CELL RESEARCH? 633
CAN WE BENEFIT FROM OTHERS' DESTRUCTION OF EMBRYOS? 633
CAN WE CREATE AN EMBRYO TO DESTROY IT? 634
SHOULD WE CLONE HUMAN EMBRYOS? 634
DOES INDUCED PLURIPOTENT STEM CELL RESEARCH ELIMINATE THE ETHICAL QUESTIONS SURROUNDING HUMAN EMBRYONIC STEM CELL RESEARCH? 635
WHAT ETHICAL GUIDELINES SHOULD GOVERN HUMAN EMBRYONIC STEM CELL, THERAPEUTIC CLONING, AND INDUCED PLURIPOTENT STEM CELL RESEARCH? 636
SUMMARY 637
KEY WORDS 637
FURTHER READING 638
Chapter 67. Stem Cell Research: Religious Considerations 640
INTRODUCTION 640
MAPPING THE TERRAIN 640
PARTICULAR TRADITIONS: AN OVERVIEW 642
ROMAN CATHOLIC CONTRIBUTIONS AND THE 14 DAY THEORY 645
KEY WORDS 648
FURTHER READING 648
Chapter 68. Stem Cell-based Therapies: Food and Drug Administration Product and Pre-Clinical Regulatory Considerations 650
ABSTRACT 650
INTRODUCTION 650
RESOURCES TO DEVELOP RECOMMENDATIONS FOR THE MANUFACTURE AND CHARACTERIZATION OF STEM CELL-BASED PRODUCTS 651
CRITICAL ELEMENTS FOR DEVELOPING A SAFE STEM CELL-BASED PRODUCT 652
GENERAL EXPECTATIONS FOR CONTROL OF THE MANUFACTURING PROCESS 657
PRECLINICAL EVALUATION SUPPORTS SAFETY ASSESSMENT: PROOF-OF-CONCEPT AND TOXICITY TESTING IN ANIMAL MODELS 657
SUMMARY 659
ACKNOWLEDGMENTS 660
KEY WORDS 660
FURTHER READING 660
Chapter 69. It's Not About Curiosity, It's About Cures: People Help Drive Progress 662
CHOOSING LIFE 662
SIZE OF THE PROMISE 662
PERSONAL PROMISES FUEL PROGRESS 663
HOPE VERSUS HYPE 664
GIVING LIFE 665
FRIENDS IN NEED AND DEED 665
PEOPLE DRIVE PROGRESS 666
BETTER HEALTH FOR ALL 667
Index 668
A 668
B 668
C 669
D 670
E 670
F 672
G 672
H 673
I 674
J 675
K 675
L 675
M 675
N 676
O 677
P 677
Q 679
R 679
S 679
T 681
U 681
V 681
W 681
X 681
Y 681
Z 681