MARC Record from marc_columbia

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020 $a8770224021$q(electronic bk.)
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035 $a(OCoLC)1269624206
037 $a9541246$bIEEE
050 4 $aQH588.S83
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049 $aZCUA
245 00 $aStem cell biology and regenerative medicine /$cedited by Charles Durand ; epilogue by Pierre Charbord.
250 $aSecond edition.
264 1 $aGistrup :$bRiver Publishers,$c2021.
300 $a1 online resource (1 volume)
336 $atext$btxt$2rdacontent
337 $acomputer$bc$2rdamedia
338 $aonline resource$bcr$2rdacarrier
490 1 $aRiver Publishers series in biotechnology and medical technology forum
520 $aThe study of stem cell biology is under intensive investigation. Because stem cells have the unique capability to self-renew and differentiate into one or several cell types, they play a critical role in development, tissue homeostasis and regeneration. Stem cells also constitute promising cell candidates for cell and gene therapy. The aim of this book is to provide readers and researchers with timely and accurate knowledge on stem cell biology and regenerative medicine. This book will cover many topics in the field and is based on conferences given by recognized scientists involved in the international master course on stem cell biology at Sorbonne Université in Paris.
588 0 $aPrint version record.
505 0 $aPreface xix List of Figures xxi List of Tables xxv List of Contributors xxvii List of Abbreviations xxxiii 1 Stem Cell Concepts 1 1.1 Introduction 1 1.2 Embryonic and Adult Stem Cells 2 1.3 The Regulation of Stem Cells and the Stem Cell Niche 9 1.4 Stem Cell Models 13 1.5 Cell Therapy Using Stem Cells 17 1.6 Concluding Remarks 20 2 Transcription Regulation by Distal Enhancers: Dynamics of the 3D Genome 33 2.1 Introduction 33 2.2 Spatio-temporal Control of Gene Activity: The Pivotal Role of Enhancers 33 2.3 The Genome-wide Enhancer Landscape 37 2.4 Enhancer Dynamics 39 2.5 Enhancers and Promoters Are Distant Structures 40 2.6 Mechanisms of Enhancer Function: The 3D Genome 41 2.7 Dynamic versus Stable Chromatin Looping 44 2.8 Transcription Factories or Foci 47 2.9 Specificity of Enhancer-Promoter Contacts 48 2.10 TADs: Topologically Associating Domains 48 2.11 Functional Relevance of TADs and Their Borders 51 2.12 Alterations of Enhancers and 3D Genome Organization in Disease 53 2.13 Concluding Remarks 55 3 Repair of DNA Double-Strand Breaks in Adult Stem Cells 71 3.1 Introduction 71 3.2 DNA Damage and Repair Mechanisms 72 3.2.1 Homologous Recombination and Related Repair Mechanisms 73 3.2.2 Classical and Alternative Non-Homologous End-Joining 74 3.2.3 Context of Repair Processes 77 3.3 Cell Response to DSBs 78 3.4 Stem Cells Resistance to Genotoxic stress 81 3.5 Efficiency and Mechanisms of DSB Repair in Adult Stem Cells 81 3.5.1 Cells in the Epithelial Tissue: Epidermal, Mammary, and Intestinal Stem Cells 81 3.5.2 Cells in the Connective Tissue: Hematopoietic and Mesenchymal Stem Cells 84 3.5.3 Skeletal Muscle Stem Cells 86 3.5.4 Neural Stem Cells 89 3.5.5 Germinal Stem Cells 89 3.5.6 Induced Pluripotent Stem Cells 90 3.6 Other Responses to DNA Damage 91 3.6.1 Apoptosis 91 3.6.2 Senescence 92 3.6.3 Differentiation 93 3.7 Concluding Remarks 94 4 Haematopoietic Stem Cell Niches in the Bone Marrow 107 4.1 Haematopoietic Stem Cell Niches 107 4.2 The Contribution of Structural Components 108 4.2.1 Vascular Niche Components 108 4.2.2 Mesenchymal Stromal Cells 109 4.2.3 Mature Haematopoietic Cells 111 4.2.4 Adipocytes 111 4.2.5 Bone-associated Cells 112 4.2.6 Innervation 112 4.3 Concluding Remarks 115 5 Computational Models of Spatio-temporal Stem Cell Organization 123 5.1 Introduction 123 5.2 Concepts of Stem Cell Organization 125 5.3 Intestinal Stem Cell Organization as a Paradigm 127 5.4 Models of Epigenetic Regulation 129 5.5 Stem Cells During Development and Aging 129 5.6 Stem Cells in Artificial Environments 132 5.7 Summary 133 5.8 Concluding Remarks 134 6 Transcriptomics Investigations 139 6.1 From Omics to Data 139 6.2 Data Analysis and Standardization in Omics 140 6.3 Study of Transcriptomes Using Microarrays 142 6.4 Study of Transcriptomes Using Next Generation Sequencing (NGS) 145 6.4.1 Steps in Transcriptome Studies Using RNA-sequencing 146 6.5 Study of Single Cell Transcriptomes 147 6.6 Network Biology 151 6.7 Machine Learning Classifiers 154 6.8 Conclusion 155 7 The Regulatory Network of X Chromosome Inactivation 163 7.1 Introduction 163 7.2 The X:Autosome Ratio Dictates Initiation of XCI 165 7.3 Initiating X-Inactivation, the Regulatory Environment of the XIC 166 7.4 The Pluripotency Network and Other Trans-acting Factors of XCI 169 7.5 The Balancing Act of Activators and Inhibitors in Initiation of XCI 171 7.6 Concluding Remarks 173 8 Directed Differentiation of Human-induced Pluripotent Stem Cells into Hepatic Cells: A Transposable Example of Disease Modeling and Regenerative Medicine Applications 181 8.1 Introduction 181 8.2 Developmental Origins of the Liver 183 8.3 hiPSCs and Directed Differentiation In Vitro 187 8.3.1 Hepatoblastic Differentiation 187 8.3.2 Hepatocyte Differentiation 187 8.3.3 Cholangiocyte Differentiation 189 8.4 hiPSC-derived Liver Cell Applications 190 8.4.1 Disease Modeling 191 8.4.2 Drug Screening and Toxicology Studies 193 8.4.3 Bio Artificial Liver Devices 194 8.4.4 Cell Transplantation 195 8.4.5 Bioprinting of 3D Liver Tissues and Whole Organ Assembly 195 8.5 Concluding Remarks 197 9 Hydra and the Evolution of Stem Cells 207 9.1 Introduction 207 9.2 The "Mystery" of Hydra's Life Cycle 208 9.3 Revisiting Stem Cells in Hydra with Emerging 212 9.4 Stem Cells in Hydra Are Controlled by Conserved 215 9.5 Epigenetic Control of the Stem Cells in Hydra 217 9.6 Taxonomically Restricted Genes (TRGs) Have Their 218 9.7 Stem Cells Interact with the Environment 220 9.8 Microbiome Contributes to Tumor Formation 224 9.9 Open Questions and Future Perspectives 226 9.10 Concluding Remarks 228 10 Regeneration in Anamniotic Vertebrates 243 10.1 Introduction 10.2 Appendage Regeneration in Amphibians 245 10.2.1 Launching Regeneration via Wounding 248 10.2.2 Role of the Nerves in Inducing Blastema 248 10.2.3 Maintenance of Blastema Outgrowth by the Shh-Fgf Activation Loop 249 10.2.4 Cellular Sources and Differentiation Potential of Blastema Cells 250 10.2.4.1 Becoming blastema cell--dedifferentiation versus stem cell activation 252 10.2.4.1.1 Muscle 252 10.2.4.1.2 Connective Tissue (CT) 253 10.2.5 Specification and Re-specification of Positional Information 254 10.2.6 Frogs--Regenerative Capacity Depends on Developmental Stage 255 10.3 Other Examples of Regeneration 256 10.3.1 Brain Regeneration 257 10.3.2 Heart Regeneration 258 10.3.3 Lens Regeneration in Amphibians via Trans Differentiation 259 10.4 Regeneration and Metabolism 261 10.5 Concluding Remarks 262 11 Stem Cells and Regeneration in Plants 273 11.1 Introduction 273 11.2 SAM Organization and Regulation 274 11.2.1 The Core WUS/CLV3 Loop 276 11.2.2 Role of Hormones in SAM Regulation 277 11.2.2.1 Cytokinin 277 11.2.2.2 Auxin 278 11.2.3 Other SAM Regulators 279 11.3 RAM Organization and Regulation 280 11.3.1 RAM Master Regulators 281 11.3.2 Role of Hormones in RAM Regulation 282 11.3.3 LR Formation 283 11.4 SAM Regeneration 284 11.4.1 Tip Regeneration 284 11.4.2 De novo Regeneration 286 11.4.3 Key Players During DNSO (de novo Shoot Organogenesis) 287 11.5 Concluding Remarks 289 11.6 Take on Messages 289 12 Hematopoietic Development in Vertebrates 297 12.1 History of the Concept of HSCs 297 12.2 On the Origin of Blood 298 12.3 An Intra-embryonic Source of HSCs 300 12.4 Construction of the Aorta and Establishment of the Dorsoventral Polarity 304 12.5 Role of the Sub-aortic Mesenchyme 306 12.6 IAHC Formation is a Conserved Mechanism in Vertebrates 309 12.7 BM ECs Can Generate a Transient Wave of HSPCs 311 12.8 Systems Used to Study Hematopoietic Cell Commitment 311 12.9 Concluding Remarks 313 13 Developmental Biology of Hematopoietic Stem Cells: Non-cell Autonomous Mechanisms 323 13.1 Introduction 323 13.2 Developmental Niches 324 13.3 Cell-intrinsic Factors of Hematopoietic Stem Cell Generation 327 13.3.1 The Endothelial-to-Hematopoietic Transition 327 13.3.2 Transcription Factor Dynamics During the EHT 328 13.4 Supporting Nascent Hematopoietic Stem Cells 331 13.4.1 Endothelial Cells 333 13.4.2 Signals From Ventral Versus Dorsal Microenvironments 334 13.4.3 Subaortic Mesenchyme 335 13.4.4 The Sympathetic Nervous System 337 13.4.5 Immune-derived Signals 337 13.4.6 Macrophages 340 13.4.7 Systemic Factors 341 13.4.7.1 Blood Flow 341 13.4.7.2 Hormone Signaling 342 13.4.7.3 Metabolism 344 13.4.8 Hypoxia 344 13.5 Future Directions 345 13.6 Conclusions 347 14 Biology of Hematopoietic Stem Cells in the Adult 359 14.1 Definition, Concepts, History 359 14.2 Characterization of HSC 360 14.2.1 Using Phenotype Analysis 360 14.2.1.1 Mouse HSC (see also4;5) 360 14.2.1.2 Human HSC 361 14.2.2 Using Functional Assays (see also 31) 363 14.2.2.1 Colony-forming cells (CFCs) and longterm culture-initiating cells (LTC-ICs) 363 14.2.2.2 Using liquid cultures 364 14.2.2.3 Using in vivo transplantation models 365 14.2.3 Physiology of HSC 366 14.2.3.1 Self-renewal and quiescence properties 366 14.2.3.2 HSC potential versus HSC fate 368 14.3 Regulation of HSC Functions 369 14.3.1 Extrinsic Regulators 369 14.3.1.1 CXCL12/CXCR4 369 14.3.1.2 Stem cell factor (SCF) and its receptor KIT 369 14.3.1.3 Integrins and Adhesion molecules 370 14.3.2 Intrinsic Regulators (see also 105) 370 14.4 Ex Vivo Expansion of HSPC 371 14.4.1 Extrinsic Factors for Ex Vivo HSPC Expansion 372 14.4.1.1 Usage of Cytokines and growth factors' combination 372 14.4.1.2 Expansion in presence of stromal cells 372 14.4.2 Developmental and Intrinsic Factors for Ex Vivo Human HSPC Expansion and or/Maintenance
650 0 $aStem cells.
650 0 $aRegenerative medicine.
650 2 $aStem Cells
650 2 $aRegenerative Medicine
650 6 $aCellules souches.
650 6 $aMédecine régénératrice.
650 7 $aRegenerative medicine.$2fast$0(OCoLC)fst01745494
650 7 $aStem cells.$2fast$0(OCoLC)fst01132975
655 4 $aElectronic books.
700 1 $aDurand, Charles,$eeditor.
776 08 $iPrint version:$tStem cell biology and regenerative medicine.$bSecond edition.$dGistrup : River Publishers, 2021$z9788770224031$w(OCoLC)1263813769
830 0 $aRiver Publishers series in biotechnology and medical technology forum.
856 40 $uhttp://www.columbia.edu/cgi-bin/cul/resolve?clio16621994$zACADEMIC - Biochemistry, Biology & Biotechnology
880 0 $6505-00/(S$aPreface xix List of Figures xxi List of Tables xxv List of Contributors xxvii List of Abbreviations xxxiii 1 Stem Cell Concepts 1 1.1 Introduction 1 1.2 Embryonic and Adult Stem Cells 2 1.3 The Regulation of Stem Cells and the Stem Cell Niche 9 1.4 Stem Cell Models 13 1.5 Cell Therapy Using Stem Cells 17 1.6 Concluding Remarks 20 2 Transcription Regulation by Distal Enhancers: Dynamics of the 3D Genome 33 2.1 Introduction 33 2.2 Spatio-temporal Control of Gene Activity: The Pivotal Role of Enhancers 33 2.3 The Genome-wide Enhancer Landscape 37 2.4 Enhancer Dynamics 39 2.5 Enhancers and Promoters Are Distant Structures 40 2.6 Mechanisms of Enhancer Function: The 3D Genome 41 2.7 Dynamic versus Stable Chromatin Looping 44 2.8 Transcription Factories or Foci 47 2.9 Specificity of Enhancer-Promoter Contacts 48 2.10 TADs: Topologically Associating Domains 48 2.11 Functional Relevance of TADs and Their Borders 51 2.12 Alterations of Enhancers and 3D Genome Organization in Disease 53 2.13 Concluding Remarks 55 3 Repair of DNA Double-Strand Breaks in Adult Stem Cells 71 3.1 Introduction 71 3.2 DNA Damage and Repair Mechanisms 72 3.2.1 Homologous Recombination and Related Repair Mechanisms 73 3.2.2 Classical and Alternative Non-Homologous End-Joining 74 3.2.3 Context of Repair Processes 77 3.3 Cell Response to DSBs 78 3.4 Stem Cells Resistance to Genotoxic stress 81 3.5 Efficiency and Mechanisms of DSB Repair in Adult Stem Cells 81 3.5.1 Cells in the Epithelial Tissue: Epidermal, Mammary, and Intestinal Stem Cells 81 3.5.2 Cells in the Connective Tissue: Hematopoietic and Mesenchymal Stem Cells 84 3.5.3 Skeletal Muscle Stem Cells 86 3.5.4 Neural Stem Cells 89 3.5.5 Germinal Stem Cells 89 3.5.6 Induced Pluripotent Stem Cells 90 3.6 Other Responses to DNA Damage 91 3.6.1 Apoptosis 91 3.6.2 Senescence 92 3.6.3 Differentiation 93 3.7 Concluding Remarks 94 4 Haematopoietic Stem Cell Niches in the Bone Marrow 107 4.1 Haematopoietic Stem Cell Niches 107 4.2 The Contribution of Structural Components 108 4.2.1 Vascular Niche Components 108 4.2.2 Mesenchymal Stromal Cells 109 4.2.3 Mature Haematopoietic Cells 111 4.2.4 Adipocytes 111 4.2.5 Bone-associated Cells 112 4.2.6 Innervation 112 4.3 Concluding Remarks 115 5 Computational Models of Spatio-temporal Stem Cell Organization 123 5.1 Introduction 123 5.2 Concepts of Stem Cell Organization 125 5.3 Intestinal Stem Cell Organization as a Paradigm 127 5.4 Models of Epigenetic Regulation 129 5.5 Stem Cells During Development and Aging 129 5.6 Stem Cells in Artificial Environments 132 5.7 Summary 133 5.8 Concluding Remarks 134 6 Transcriptomics Investigations 139 6.1 From Omics to Data 139 6.2 Data Analysis and Standardization in Omics 140 6.3 Study of Transcriptomes Using Microarrays 142 6.4 Study of Transcriptomes Using Next Generation Sequencing (NGS) 145 6.4.1 Steps in Transcriptome Studies Using RNA-sequencing 146 6.5 Study of Single Cell Transcriptomes 147 6.6 Network Biology 151 6.7 Machine Learning Classifiers 154 6.8 Conclusion 155 7 The Regulatory Network of X Chromosome Inactivation 163 7.1 Introduction 163 7.2 The X:Autosome Ratio Dictates Initiation of XCI 165 7.3 Initiating X-Inactivation, the Regulatory Environment of the XIC 166 7.4 The Pluripotency Network and Other Trans-acting Factors of XCI 169 7.5 The Balancing Act of Activators and Inhibitors in Initiation of XCI 171 7.6 Concluding Remarks 173 8 Directed Differentiation of Human-induced Pluripotent Stem Cells into Hepatic Cells: A Transposable Example of Disease Modeling and Regenerative Medicine Applications 181 8.1 Introduction 181 8.2 Developmental Origins of the Liver 183 8.3 hiPSCs and Directed Differentiation In Vitro 187 8.3.1 Hepatoblastic Differentiation 187 8.3.2 Hepatocyte Differentiation 187 8.3.3 Cholangiocyte Differentiation 189 8.4 hiPSC-derived Liver Cell Applications 190 8.4.1 Disease Modeling 191 8.4.2 Drug Screening and Toxicology Studies 193 8.4.3 Bio Artificial Liver Devices 194 8.4.4 Cell Transplantation 195 8.4.5 Bioprinting of 3D Liver Tissues and Whole Organ Assembly 195 8.5 Concluding Remarks 197 9 Hydra and the Evolution of Stem Cells 207 9.1 Introduction 207 9.2 The "Mystery" of Hydra's Life Cycle 208 9.3 Revisiting Stem Cells in Hydra with Emerging 212 9.4 Stem Cells in Hydra Are Controlled by Conserved 215 9.5 Epigenetic Control of the Stem Cells in Hydra 217 9.6 Taxonomically Restricted Genes (TRGs) Have Their 218 9.7 Stem Cells Interact with the Environment 220 9.8 Microbiome Contributes to Tumor Formation 224 9.9 Open Questions and Future Perspectives 226 9.10 Concluding Remarks 228 10 Regeneration in Anamniotic Vertebrates 243 10.1 Introduction 10.2 Appendage Regeneration in Amphibians 245 10.2.1 Launching Regeneration via Wounding 248 10.2.2 Role of the Nerves in Inducing Blastema 248 10.2.3 Maintenance of Blastema Outgrowth by the Shh-Fgf Activation Loop 249 10.2.4 Cellular Sources and Differentiation Potential of Blastema Cells 250 10.2.4.1 Becoming blastema cell--dedifferentiation versus stem cell activation 252 10.2.4.1.1 Muscle 252 10.2.4.1.2 Connective Tissue (CT) 253 10.2.5 Specification and Re-specification of Positional Information 254 10.2.6 Frogs--Regenerative Capacity Depends on Developmental Stage 255 10.3 Other Examples of Regeneration 256 10.3.1 Brain Regeneration 257 10.3.2 Heart Regeneration 258 10.3.3 Lens Regeneration in Amphibians via Trans Differentiation 259 10.4 Regeneration and Metabolism 261 10.5 Concluding Remarks 262 11 Stem Cells and Regeneration in Plants 273 11.1 Introduction 273 11.2 SAM Organization and Regulation 274 11.2.1 The Core WUS/CLV3 Loop 276 11.2.2 Role of Hormones in SAM Regulation 277 11.2.2.1 Cytokinin 277 11.2.2.2 Auxin 278 11.2.3 Other SAM Regulators 279 11.3 RAM Organization and Regulation 280 11.3.1 RAM Master Regulators 281 11.3.2 Role of Hormones in RAM Regulation 282 11.3.3 LR Formation 283 11.4 SAM Regeneration 284 11.4.1 Tip Regeneration 284 11.4.2 De novo Regeneration 286 11.4.3 Key Players During DNSO (de novo Shoot Organogenesis) 287 11.5 Concluding Remarks 289 11.6 Take on Messages 289 12 Hematopoietic Development in Vertebrates 297 12.1 History of the Concept of HSCs 297 12.2 On the Origin of Blood 298 12.3 An Intra-embryonic Source of HSCs 300 12.4 Construction of the Aorta and Establishment of the Dorsoventral Polarity 304 12.5 Role of the Sub-aortic Mesenchyme 306 12.6 IAHC Formation is a Conserved Mechanism in Vertebrates 309 12.7 BM ECs Can Generate a Transient Wave of HSPCs 311 12.8 Systems Used to Study Hematopoietic Cell Commitment 311 12.9 Concluding Remarks 313 13 Developmental Biology of Hematopoietic Stem Cells: Non-cell Autonomous Mechanisms 323 13.1 Introduction 323 13.2 Developmental Niches 324 13.3 Cell-intrinsic Factors of Hematopoietic Stem Cell Generation 327 13.3.1 The Endothelial-to-Hematopoietic Transition 327 13.3.2 Transcription Factor Dynamics During the EHT 328 13.4 Supporting Nascent Hematopoietic Stem Cells 331 13.4.1 Endothelial Cells 333 13.4.2 Signals From Ventral Versus Dorsal Microenvironments 334 13.4.3 Subaortic Mesenchyme 335 13.4.4 The Sympathetic Nervous System 337 13.4.5 Immune-derived Signals 337 13.4.6 Macrophages 340 13.4.7 Systemic Factors 341 13.4.7.1 Blood Flow 341 13.4.7.2 Hormone Signaling 342 13.4.7.3 Metabolism 344 13.4.8 Hypoxia 344 13.5 Future Directions 345 13.6 Conclusions 347 14 Biology of Hematopoietic Stem Cells in the Adult 359 14.1 Definition, Concepts, History 359 14.2 Characterization of HSC 360 14.2.1 Using Phenotype Analysis 360 14.2.1.1 Mouse HSC (see also4;5) 360 14.2.1.2 Human HSC 361 14.2.2 Using Functional Assays (see also 31) 363 14.2.2.1 Colony-forming cells (CFCs) and longterm culture-initiating cells (LTC-ICs) 363 14.2.2.2 Using liquid cultures 364 14.2.2.3 Using in vivo transplantation models 365 14.2.3 Physiology of HSC 366 14.2.3.1 Self-renewal and quiescence properties 366 14.2.3.2 HSC potential versus HSC fate 368 14.3 Regulation of HSC Functions 369 14.3.1 Extrinsic Regulators 369 14.3.1.1 CXCL12/CXCR4 369 14.3.1.2 Stem cell factor (SCF) and its receptor KIT 369 14.3.1.3 Integrins and Adhesion molecules 370 14.3.2 Intrinsic Regulators (see also 105) 370 14.4 Ex Vivo Expansion of HSPC 371 14.4.1 Extrinsic Factors for Ex Vivo HSPC Expansion 372 14.4.1.1 Usage of Cytokines and growth factors' combination 372 14.4.1.2 Expansion in presence of stromal cells 372 14.4.2 Developmental and Intrinsic Factors for Ex Vivo Human HSPC Expansion and or/Maintenance 373 14.4.2.1 Approaches Using Delivery of Intrinsic Factors 373 14.4.2.2 A. HOXB4-mediated expansion 373 14.4.2.3 B. Other potential molecular targets (Notch and Wnt pathways) 373 14.4.2.4 Approaches using modification of human HSPC microenvironment: Focus on the Hypoxia/HIF pathway 374 14.4.3 Chemical Compounds in the Era of Advanced Technologies for Ex Vivo Human HSPC Expansion 376 14.5 Conclusive Remarks and Perspectives 377 15 Epithelial Stem Cells in the Skin 393 15.1 Introduction 393 15.2 The Interfollicular Epidermis 395 15.2.1 Development of the IFE 395 15.2.2 IFE during Postnatal Growth 396 15.2.3 IFE in Adulthood 397 15.3 The Hair Follicle 399 15.3.1 HF Development 400 15.3.2 HF Cycling 401 15.4 Contribution of Bulge Stem Cells to the Epidermis 402 15.5 Contribution of Bulge Stem Cells to the Sebaceous Gland Lineage 404 15.6 The Sweat Gland 406 15.7 Concluding Remarks 407 16 Mammary Stem Cells 415 16.1 Introduction 415 16.2 The Mammary Epithelium and Its Stem Cells 416 16.3 Lineage Tracing Analysis to Study Stem Cells In Vivo and In Situ 418 16.4 Mammary Cell Plasticity 420 16.5 Molecular Signals Governing MaSCs and Gland Morphogenesis 422 16.5.1 The Notch Pathway 422 16.5.2 The Wnt Pathway 425 16.5.3 The FGF pathway 426 16.5.4 The TGF β Pathway 426 16.5.5 The Hedgehog Pathway 427 16.6 Concludi ...
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