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简介
Principles of Cloning is the first comprehensive book on animal cloning since the creation of Dolly. The contributing authors are the principal investigators on each of the animal species cloned to date, and are expertly qualified to present the state-of-the-art information in their respective areas. Editors Cibelli, Lanza and West garnered worldwide spotlight late in 2001 when their company, Advanced Cell Technology, announced the successful engineering of the world's first cloned human embryo. The trio was featured in the US News & WorldReport December 2001 cover story, "The First Human Clone."The book presents the basic biological mechanisms of how cloning works and progresses to discuss current and potential applications in basic biology, agriculture, biotechnology, and medicine. Key Features * First and mostcomprehensive book on animal cloning * Chapters written bythe world' expert in each area * From the early experiments in amphibia to the latest one in mammals, everything is included in this book and told by the researcher that did it and how they did it * Basic biological mechanisms on how cloning works and all their current and potential applications * Cloning applications on basic biology, agriculture, biotechnology and medicine are included * Editors are the pioneers in the field.
目录
Front Cover 1
Principles of Cloning 4
CONTENTS 8
CONTRIBUTORS 16
PREFACE 20
FOREWORD 22
Introduction 24
Chapter 1. Historical Perspective 26
ANCIENT CLONING (ASEXUAL REPRODUCTION)\u2014IN THE BEGINNING 26
SPONTANEOUS REGENERATION 27
TYPES OF CLONING 27
EXAMPLES OF NATURAL CLONING IN HUMANS AND ANIMALS 27
EXPERIMENTAL CLONING OF NONMAMMALS 27
CLONING MAMMALS 29
REFERENCES 37
Part I: Basic Biological Processes 42
Chapter 2. Activation of Mammalian Oocytes 44
INTRODUCTION 44
REQUIREMENT OF Ca2+ FOR OOCYTE ACTIVATION 45
HOW DOES THE SPERM TRIGGER Ca2+ RELEASE? 48
REGULATION OF FERTILIZATION-ASSOCIATED [Ca2+]i OSCILLATIONS 53
REQUIREMENTS FOR ASSEMBLY OF COMPETENT PRONUCLEI 55
[Ca2+]i OSCILLATIONS AND APOPTOSIS IN MAMMALIAN OOCYTES 60
REFERENCES 62
Chapter 3. The Nucleus 70
DNA AND GENOME STRUCTURE 71
DNA-ASSOCIATED PROTEINS IN SOMATIC AND GERM CELLS 72
THE NUCLEAR ENVELOPE, LAMINA, AND MATRIX: E PLURIBUS UNUM 92
PERSPECTIVE 96
UPDATE 96
REFERENCES 96
Chapter 4. Nuclear Reprogramming: Biological and Technological Constraints 108
NUCLEAR TRANSFER TECHNOLOGY AND ITS INFLUENCE ON THE DEVELOPMENT OF CLONED MAMMALS 108
EPIGENETIC REPROGRAMMING AFTER NUCLEAR TRANSFER 114
REFERENCES 119
Chapter 5. Plasticity of Somatic Nucleus by Epigenetic Reprogramming Via Cell Hybridization 122
INTRODUCTION 122
NUCLEAR REPROGRAMMING OF SOMATIC CELLS BY HYBRIDIZATION WITH EMBRYONIC STEM CELLS 123
NUCLEAR REPROGRAMMING OF SOMATIC CELLS BY HYBRIDIZATION WITH EMBRYONIC GERM CELLS 127
CLONING BY SOMATIC CELL TRANSPLANTATION INTO OOCYTES 129
CONCLUSION 129
REFERENCES 129
Chapter 6. Determinants of Pluripotency in Mammals 132
INTRODUCTION 132
DEFINING PLURIPOTENCY BY MEANS OF TOTIPOTENCY 133
WHY PLURIPOTENCY? 135
THE FIELDS OF PLURIPOTENCY 136
LEVELS OF CELL POTENCY 139
THE CELLULAR BASES OF POTENCY 142
PLURIPOTENTIAL STEM CELLS AND CLONING 147
METHODS OF ADDRESSING CELL POTENCY 148
MARKERS OF PLURIPOTENCY 157
OCT4, A MODERN TOOL FOR ANALYSIS OF PLURIPOTENCY 158
LESSONS FROM CLONING 167
REFERENCES 168
Part II: Methods 176
Chapter 7. Micromanipulation Techniques for Cloning 178
INTRODUCTION 178
MAKING MANIPULATION TOOLS 178
MICROSCOPY AND EQUIPMENT FOR MICROMANIPULATION 183
MICROMANIPULATION PROCEDURES 186
PIEZOELECTRIC ASSISTED NUCLEAR TRANSFER 192
TECHNICAL IMPROVEMENTS 194
REFERENCES 195
Chapter 8. Microinsemination and Nuclear Transfer with Male Germ Cells 198
INTRODUCTION 198
MICROINSEMINATION 200
NUCLEAR TRANSFER WITH MALE PRIMORDIAL GERM CELLS 205
REFERENCES 206
Chapter 9. Development of Viable Mammalian Embryos in Vitro: Evolution of Sequential Media 210
INTRODUCTION 210
DYNAMICS OF EMBRYO AND MATERNAL PHYSIOLOGY 210
METABOLISM OF THE EMBRYO 215
DECREASING INTRACELLULAR STRESS 218
EVOLUTION OF SEQUENTIAL EMBRYO CULTURE MEDIA 218
FACTORS OTHER THAN MEDIUM FORMULATION THAT IMPACT EMBRYO DEVELOPMENT AND VIABILITY 222
INHERENT PROBLEMS OF COCULTURE 229
CONCLUSIONS 231
REFERENCES 231
Chapter 10. Genetic and Phenotypic Similarity Among Members of Mammalian Clonal Sets 238
DEFINITION OF CLONING 238
CYTOPLASMIC GENETICS 239
EPIGENETIC EFFECTS 239
UTERINE EFFECTS 241
NEONATAL ENVIRONMENT 242
LARGE-OFFSPRING SYNDROME 242
MUTATIONS 243
CULTURAL INHERITANCE 244
HOW SIMILAR? 244
SUMMARY AND PERSPECTIVE 246
REFERENCES 246
Chapter 11. Genetic Modification and Cloning in Mammals 250
GENERAL INTRODUCTION 250
HISTORICAL PERSPECTIVE 250
CONVENTIONAL ENRICHMENT PROTOCOLS TO ENHANCE HOMOLOGOUS RECOMBINATION 252
MODULATION OF HOMOLOGOUS RECOMBINATION AND NONHOMOLOGOUS END JOINING DNA REPAIR PATHWAYS 253
GENE TARGETING: EXPLOITING MECHANISMS OF DNA DOUBLE-STRAND BREAK REPAIR IN MAMMALIAN CELLS 254
MODELS OF HOMOLOGOUS RECOMBINATION REPAIR 254
BIOCHEMISTRY OF HR REPAIR 255
OVEREXPRESSION OF NORMAL AND DOMINANT-NEGATIVE FORMS OF HR PROTEINS ENHANCE HOMOLOGOUS RECOMBINATION 257
RANDOM INSERTION OF TARGETING CONSTRUCTS MEDIATED BY NONHOMOLOGOUS END JOINING 260
SUPPRESSING NONHOMOLOGOUS END JOINING PATHWAY BY INHIBITING KEY NHEJ PROTEINS 261
REPLICATIVE SENESCENCE 261
RELATIONSHIP BETWEEN SENESCENCE AND DNA REPAIR 263
SUMMARY 263
REFERENCES 264
Chapter 12. Pregnancy and Neonatal Care of Cloned Animals 270
OCCURRENCE OF LARGE-OFFSPRING SYNDROME 270
GESTATIONAL MONITORING 272
PERINATAL MANAGEMENT 280
SUMMARY 285
REFERENCES 285
Chapter 13. Donor Cell Type and Cloning Efficiency in Mammals 290
DEVELOPMENTAL POTENTIAL OF GERM LINE NUCLEI AT VARIOUS CELL CYCLE STAGES 290
DEVELOPMENTAL POTENTIAL OF SOMATIC CELL LINE NUCLEI FROM DIFFERENT TISSUES 291
REFERENCES 298
Part III: Cloning by Species 302
Chapter 14. Cloning of Amphibians 304
INTRODUCTION 304
EGG FORMATION AND EMBRYO DEVELOPMENT 305
METHODOLOGY 306
NUCLEAR TRANSFER SUCCESS 307
NUCLEAR REPROGRAMMING 308
REFERENCES 309
Chapter 15. Cloning of Fish 310
INTRODUCTION 310
DEVELOPMENT OF BASIC TECHNIQUES OF TRANSPLANTATION OF EMBRYONIC CELLS TO NONENUCLEATED EGGS 311
EXPRESSION OF FOREIGN GENES IN NUCLEAR TRANSPLANTS 315
FERTILE AND DIPLOID NUCLEAR TRANSPLANTS 317
CONCLUSIONS AND PERSPECTIVES 320
REFERENCES 320
Chapter 16. Cloning of Mice 324
INTRODUCTION 324
CONTRIBUTION OF THE RECIPIENT CYTOPLASM 328
CONTRIBUTION OF THE NUCLEUS DONOR 331
TECHNICAL DETERMINANTS OF DEVELOPMENTAL OUTCOME IN CLONING 341
CLONE DEVELOPMENT 345
QUESTIONS AND APPLICATIONS 353
CONCLUDING REMARKS 355
REFERENCES 359
Chapter 17. Cloning of Rabbits 366
INTRODUCTION 366
BACKGROUND OF RABBIT EMBRYOLOGY 367
STATE OF THE ART 369
APPLICATIONS OF SOMATIC CELL NUCLEAR TRANSFER IN RABBITS 379
APPENDIX: PROTOCOL FOR RABBIT SOMATIC-CELL NUCLEAR TRANSFER 379
REFERENCES 384
Chapter 18. Nuclear Transfer in Swine 390
INTRODUCTION 390
INITIAL STUDIES 390
MOLECULAR EVENTS OF REPROGRAMMING AS REVEALED BY STUDIES IN PIGS 391
OOCYTE ACTIVATION 393
IN VITRO DEVELOPMENT OF NUCLEAR TRANSFER EMBRYOS 393
PRODUCTION OF OFFSPRING RESULTING FROM NUCLEAR TRANSFER 394
CONCLUSIONS 395
REFERENCES 395
Chapter 19. Cloning of cattle 398
INTRODUCTION 398
HISTORY 398
CLONING FROM FETAL CELLS 400
CLONING FROM ADULT SOMATIC CELLS 402
TELOMERES 404
LARGE-OFFSPRING SYNDROME 405
APPLICATIONS OF NUCLEAR TRANSFER 408
REFERENCES 410
Chapter 20. Cell Cycle Regulation in Cloning 414
INTRODUCTION 414
INITIAL EVENTS 414
OOCYTE DEVELOPMENT AND THE CELL CYCLE 415
CELL CYCLE EFFECTS OF OOCYTE-DERIVED CYTOPLAST RECIPIENTS 416
OTHER FACTORS RELATED TO THE RECIPIENT CELL CYCLE PHASE 418
EFFECTS OF CELL CYCLE COMBINATIONS ON DEVELOPMENT 419
EFFECTS OF THE DONOR CELL CYCLE STAGE ON DEVELOPMENT 420
SUMMARY 420
REFERENCES 420
Part IV: Currently Sought after Species 424
Chapter 21. Cloning of Rats 426
INTRODUCTION 426
WHY CLONE THE RAT? 426
STEPS IN CLONING 427
DONOR CELLS 431
ACTIVATION 432
OUTCOMES 433
TRANSFER OF RECONSTITUTED EMBRYOS TO SURROGATE MOTHERS 433
LIMITATIONS 434
QUESTIONS OF INTEREST IN CLONING 436
REFERENCES 437
Part V: Nuclear Transfer in Primates 440
Chapter 22. Cloning in Nonhuman Primates 442
INTRODUCTION 442
WHY GENETICALLY IDENTICAL NONHUMAN PRIMATES? 442
FACTORS INFLUENCING RESEARCH IN ASSISTED REPRODUCTIVE TECHNOLOGIES 443
METHODS FOR PRODUCTION OF GENETICALLY IDENTICAL ANIMALS 445
CONCLUSIONS 451
REFERENCES 452
Part VI: Applications 456
Chapter 23. Nuclear Transfer for Stem Cells 458
INTRODUCTION 458
NUCLEAR TRANSFER USING SOMATIC CELLS TO PRODUCE ES CELLS 458
STEM CELL HYBRIDIZATION 462
CONCLUSIONS 463
REFERENCES 463
Chapter 24. Current Research and Commercial Applications of Cloning Technology 466
INTRODUCTION 466
CLONING RESEARCH ANIMALS 467
PREVIOUS AND PRESENT OBSTACLES TO COMMERCIAL CLONING IN ANIMAL AGRICULTURE 471
SUMMARY 478
REFERENCES 478
Chapter 25. Transgenic Cloned Goats and the Production of Therapeutic Proteins 482
INTRODUCTION 482
MAMMARY GLAND-SPECIFIC TRANSGENES FOR SOMATIC CELL NUCLEAR TRANSFER 485
GENERATION OF TRANSGENIC FOUNDERS BY TRANSFECTED SOMATIC CELL NUCLEAR TRANSFER 486
PRODUCTION OF TRANSGENIC GOATS BY ADULT SOMATIC CELL NUCLEAR TRANSFER 492
IN VITRO-DERIVED OOCYTES VS. IN VIVO-DERIVED OOCYTES 493
CONCLUSION 494
REFERENCES 495
Part VII: Ethical and Legal Affairs 498
Chapter 26. Ethical Implications of Cloning 500
INTRODUCTION 500
REPRODUCTIVE CLONING 501
THERAPEUTIC CLONING 510
CONCLUSION 514
REFERENCES 514
Final Remarks 518
Chapter 27. Mammalian Cloning: Challenges for the Future 520
INTRODUCTION 520
ACTIVATION OF DEVELOPMENT 520
THERAPEUTIC CLONING VERSUS DIRECT REPROGRAMMING OF ADULT CELLS 521
REPRODUCTIVE CLONING\u2014QUESTIONS, MYTHS, AND CONCERNS 522
REFERENCES 523
AUTHOR INDEX 526
SUBJECT INDEX 546
Principles of Cloning 4
CONTENTS 8
CONTRIBUTORS 16
PREFACE 20
FOREWORD 22
Introduction 24
Chapter 1. Historical Perspective 26
ANCIENT CLONING (ASEXUAL REPRODUCTION)\u2014IN THE BEGINNING 26
SPONTANEOUS REGENERATION 27
TYPES OF CLONING 27
EXAMPLES OF NATURAL CLONING IN HUMANS AND ANIMALS 27
EXPERIMENTAL CLONING OF NONMAMMALS 27
CLONING MAMMALS 29
REFERENCES 37
Part I: Basic Biological Processes 42
Chapter 2. Activation of Mammalian Oocytes 44
INTRODUCTION 44
REQUIREMENT OF Ca2+ FOR OOCYTE ACTIVATION 45
HOW DOES THE SPERM TRIGGER Ca2+ RELEASE? 48
REGULATION OF FERTILIZATION-ASSOCIATED [Ca2+]i OSCILLATIONS 53
REQUIREMENTS FOR ASSEMBLY OF COMPETENT PRONUCLEI 55
[Ca2+]i OSCILLATIONS AND APOPTOSIS IN MAMMALIAN OOCYTES 60
REFERENCES 62
Chapter 3. The Nucleus 70
DNA AND GENOME STRUCTURE 71
DNA-ASSOCIATED PROTEINS IN SOMATIC AND GERM CELLS 72
THE NUCLEAR ENVELOPE, LAMINA, AND MATRIX: E PLURIBUS UNUM 92
PERSPECTIVE 96
UPDATE 96
REFERENCES 96
Chapter 4. Nuclear Reprogramming: Biological and Technological Constraints 108
NUCLEAR TRANSFER TECHNOLOGY AND ITS INFLUENCE ON THE DEVELOPMENT OF CLONED MAMMALS 108
EPIGENETIC REPROGRAMMING AFTER NUCLEAR TRANSFER 114
REFERENCES 119
Chapter 5. Plasticity of Somatic Nucleus by Epigenetic Reprogramming Via Cell Hybridization 122
INTRODUCTION 122
NUCLEAR REPROGRAMMING OF SOMATIC CELLS BY HYBRIDIZATION WITH EMBRYONIC STEM CELLS 123
NUCLEAR REPROGRAMMING OF SOMATIC CELLS BY HYBRIDIZATION WITH EMBRYONIC GERM CELLS 127
CLONING BY SOMATIC CELL TRANSPLANTATION INTO OOCYTES 129
CONCLUSION 129
REFERENCES 129
Chapter 6. Determinants of Pluripotency in Mammals 132
INTRODUCTION 132
DEFINING PLURIPOTENCY BY MEANS OF TOTIPOTENCY 133
WHY PLURIPOTENCY? 135
THE FIELDS OF PLURIPOTENCY 136
LEVELS OF CELL POTENCY 139
THE CELLULAR BASES OF POTENCY 142
PLURIPOTENTIAL STEM CELLS AND CLONING 147
METHODS OF ADDRESSING CELL POTENCY 148
MARKERS OF PLURIPOTENCY 157
OCT4, A MODERN TOOL FOR ANALYSIS OF PLURIPOTENCY 158
LESSONS FROM CLONING 167
REFERENCES 168
Part II: Methods 176
Chapter 7. Micromanipulation Techniques for Cloning 178
INTRODUCTION 178
MAKING MANIPULATION TOOLS 178
MICROSCOPY AND EQUIPMENT FOR MICROMANIPULATION 183
MICROMANIPULATION PROCEDURES 186
PIEZOELECTRIC ASSISTED NUCLEAR TRANSFER 192
TECHNICAL IMPROVEMENTS 194
REFERENCES 195
Chapter 8. Microinsemination and Nuclear Transfer with Male Germ Cells 198
INTRODUCTION 198
MICROINSEMINATION 200
NUCLEAR TRANSFER WITH MALE PRIMORDIAL GERM CELLS 205
REFERENCES 206
Chapter 9. Development of Viable Mammalian Embryos in Vitro: Evolution of Sequential Media 210
INTRODUCTION 210
DYNAMICS OF EMBRYO AND MATERNAL PHYSIOLOGY 210
METABOLISM OF THE EMBRYO 215
DECREASING INTRACELLULAR STRESS 218
EVOLUTION OF SEQUENTIAL EMBRYO CULTURE MEDIA 218
FACTORS OTHER THAN MEDIUM FORMULATION THAT IMPACT EMBRYO DEVELOPMENT AND VIABILITY 222
INHERENT PROBLEMS OF COCULTURE 229
CONCLUSIONS 231
REFERENCES 231
Chapter 10. Genetic and Phenotypic Similarity Among Members of Mammalian Clonal Sets 238
DEFINITION OF CLONING 238
CYTOPLASMIC GENETICS 239
EPIGENETIC EFFECTS 239
UTERINE EFFECTS 241
NEONATAL ENVIRONMENT 242
LARGE-OFFSPRING SYNDROME 242
MUTATIONS 243
CULTURAL INHERITANCE 244
HOW SIMILAR? 244
SUMMARY AND PERSPECTIVE 246
REFERENCES 246
Chapter 11. Genetic Modification and Cloning in Mammals 250
GENERAL INTRODUCTION 250
HISTORICAL PERSPECTIVE 250
CONVENTIONAL ENRICHMENT PROTOCOLS TO ENHANCE HOMOLOGOUS RECOMBINATION 252
MODULATION OF HOMOLOGOUS RECOMBINATION AND NONHOMOLOGOUS END JOINING DNA REPAIR PATHWAYS 253
GENE TARGETING: EXPLOITING MECHANISMS OF DNA DOUBLE-STRAND BREAK REPAIR IN MAMMALIAN CELLS 254
MODELS OF HOMOLOGOUS RECOMBINATION REPAIR 254
BIOCHEMISTRY OF HR REPAIR 255
OVEREXPRESSION OF NORMAL AND DOMINANT-NEGATIVE FORMS OF HR PROTEINS ENHANCE HOMOLOGOUS RECOMBINATION 257
RANDOM INSERTION OF TARGETING CONSTRUCTS MEDIATED BY NONHOMOLOGOUS END JOINING 260
SUPPRESSING NONHOMOLOGOUS END JOINING PATHWAY BY INHIBITING KEY NHEJ PROTEINS 261
REPLICATIVE SENESCENCE 261
RELATIONSHIP BETWEEN SENESCENCE AND DNA REPAIR 263
SUMMARY 263
REFERENCES 264
Chapter 12. Pregnancy and Neonatal Care of Cloned Animals 270
OCCURRENCE OF LARGE-OFFSPRING SYNDROME 270
GESTATIONAL MONITORING 272
PERINATAL MANAGEMENT 280
SUMMARY 285
REFERENCES 285
Chapter 13. Donor Cell Type and Cloning Efficiency in Mammals 290
DEVELOPMENTAL POTENTIAL OF GERM LINE NUCLEI AT VARIOUS CELL CYCLE STAGES 290
DEVELOPMENTAL POTENTIAL OF SOMATIC CELL LINE NUCLEI FROM DIFFERENT TISSUES 291
REFERENCES 298
Part III: Cloning by Species 302
Chapter 14. Cloning of Amphibians 304
INTRODUCTION 304
EGG FORMATION AND EMBRYO DEVELOPMENT 305
METHODOLOGY 306
NUCLEAR TRANSFER SUCCESS 307
NUCLEAR REPROGRAMMING 308
REFERENCES 309
Chapter 15. Cloning of Fish 310
INTRODUCTION 310
DEVELOPMENT OF BASIC TECHNIQUES OF TRANSPLANTATION OF EMBRYONIC CELLS TO NONENUCLEATED EGGS 311
EXPRESSION OF FOREIGN GENES IN NUCLEAR TRANSPLANTS 315
FERTILE AND DIPLOID NUCLEAR TRANSPLANTS 317
CONCLUSIONS AND PERSPECTIVES 320
REFERENCES 320
Chapter 16. Cloning of Mice 324
INTRODUCTION 324
CONTRIBUTION OF THE RECIPIENT CYTOPLASM 328
CONTRIBUTION OF THE NUCLEUS DONOR 331
TECHNICAL DETERMINANTS OF DEVELOPMENTAL OUTCOME IN CLONING 341
CLONE DEVELOPMENT 345
QUESTIONS AND APPLICATIONS 353
CONCLUDING REMARKS 355
REFERENCES 359
Chapter 17. Cloning of Rabbits 366
INTRODUCTION 366
BACKGROUND OF RABBIT EMBRYOLOGY 367
STATE OF THE ART 369
APPLICATIONS OF SOMATIC CELL NUCLEAR TRANSFER IN RABBITS 379
APPENDIX: PROTOCOL FOR RABBIT SOMATIC-CELL NUCLEAR TRANSFER 379
REFERENCES 384
Chapter 18. Nuclear Transfer in Swine 390
INTRODUCTION 390
INITIAL STUDIES 390
MOLECULAR EVENTS OF REPROGRAMMING AS REVEALED BY STUDIES IN PIGS 391
OOCYTE ACTIVATION 393
IN VITRO DEVELOPMENT OF NUCLEAR TRANSFER EMBRYOS 393
PRODUCTION OF OFFSPRING RESULTING FROM NUCLEAR TRANSFER 394
CONCLUSIONS 395
REFERENCES 395
Chapter 19. Cloning of cattle 398
INTRODUCTION 398
HISTORY 398
CLONING FROM FETAL CELLS 400
CLONING FROM ADULT SOMATIC CELLS 402
TELOMERES 404
LARGE-OFFSPRING SYNDROME 405
APPLICATIONS OF NUCLEAR TRANSFER 408
REFERENCES 410
Chapter 20. Cell Cycle Regulation in Cloning 414
INTRODUCTION 414
INITIAL EVENTS 414
OOCYTE DEVELOPMENT AND THE CELL CYCLE 415
CELL CYCLE EFFECTS OF OOCYTE-DERIVED CYTOPLAST RECIPIENTS 416
OTHER FACTORS RELATED TO THE RECIPIENT CELL CYCLE PHASE 418
EFFECTS OF CELL CYCLE COMBINATIONS ON DEVELOPMENT 419
EFFECTS OF THE DONOR CELL CYCLE STAGE ON DEVELOPMENT 420
SUMMARY 420
REFERENCES 420
Part IV: Currently Sought after Species 424
Chapter 21. Cloning of Rats 426
INTRODUCTION 426
WHY CLONE THE RAT? 426
STEPS IN CLONING 427
DONOR CELLS 431
ACTIVATION 432
OUTCOMES 433
TRANSFER OF RECONSTITUTED EMBRYOS TO SURROGATE MOTHERS 433
LIMITATIONS 434
QUESTIONS OF INTEREST IN CLONING 436
REFERENCES 437
Part V: Nuclear Transfer in Primates 440
Chapter 22. Cloning in Nonhuman Primates 442
INTRODUCTION 442
WHY GENETICALLY IDENTICAL NONHUMAN PRIMATES? 442
FACTORS INFLUENCING RESEARCH IN ASSISTED REPRODUCTIVE TECHNOLOGIES 443
METHODS FOR PRODUCTION OF GENETICALLY IDENTICAL ANIMALS 445
CONCLUSIONS 451
REFERENCES 452
Part VI: Applications 456
Chapter 23. Nuclear Transfer for Stem Cells 458
INTRODUCTION 458
NUCLEAR TRANSFER USING SOMATIC CELLS TO PRODUCE ES CELLS 458
STEM CELL HYBRIDIZATION 462
CONCLUSIONS 463
REFERENCES 463
Chapter 24. Current Research and Commercial Applications of Cloning Technology 466
INTRODUCTION 466
CLONING RESEARCH ANIMALS 467
PREVIOUS AND PRESENT OBSTACLES TO COMMERCIAL CLONING IN ANIMAL AGRICULTURE 471
SUMMARY 478
REFERENCES 478
Chapter 25. Transgenic Cloned Goats and the Production of Therapeutic Proteins 482
INTRODUCTION 482
MAMMARY GLAND-SPECIFIC TRANSGENES FOR SOMATIC CELL NUCLEAR TRANSFER 485
GENERATION OF TRANSGENIC FOUNDERS BY TRANSFECTED SOMATIC CELL NUCLEAR TRANSFER 486
PRODUCTION OF TRANSGENIC GOATS BY ADULT SOMATIC CELL NUCLEAR TRANSFER 492
IN VITRO-DERIVED OOCYTES VS. IN VIVO-DERIVED OOCYTES 493
CONCLUSION 494
REFERENCES 495
Part VII: Ethical and Legal Affairs 498
Chapter 26. Ethical Implications of Cloning 500
INTRODUCTION 500
REPRODUCTIVE CLONING 501
THERAPEUTIC CLONING 510
CONCLUSION 514
REFERENCES 514
Final Remarks 518
Chapter 27. Mammalian Cloning: Challenges for the Future 520
INTRODUCTION 520
ACTIVATION OF DEVELOPMENT 520
THERAPEUTIC CLONING VERSUS DIRECT REPROGRAMMING OF ADULT CELLS 521
REPRODUCTIVE CLONING\u2014QUESTIONS, MYTHS, AND CONCERNS 522
REFERENCES 523
AUTHOR INDEX 526
SUBJECT INDEX 546
Principles of cloning /
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