Preface to the Professor xvii
Preface to the Student xxi
Acknowledgments xxiii
List of Figures xxv
List of Tables xxxvii
List of Case Studies xxxix
List of Focus Boxes xli
1 A HISTORY OF STEM CELL RESEARCH 1
Early Studies 1
Hematopoietic Stem Cell Discovery 4
Mouse Embryonic Stem Cell Discovery 6
Successful Neural Stem Cell Culture 7
The Discovery of Cancer Stem Cells 8
Human Embryonic Stem Cell Discovery 9
Stem Cells And Cloning 11
Cord Blood Embryonic]Like Stem Cells—An Alternative to Es and Adult Stem Cells 14
Breakthrough In Spinal Cord Injury Repair 15
The Generation of ips Cells 16
iPS Cells Derived from Keratinocytes 20
iPS Induction Without the Use of Viruses 20
Transposon]Mediated iPS 21
Protein]Based iPS 22
The Discovery of Human Amniotic Stem Cells 24
Human Embryonic Stem Cells Generated Without Embryo Destruction 25
Human Cloning 25
Mesenchymal Stem Cell]Derived Human Knee Cartilage 27
The First Clinical Trial Using Human Embryonic Stem Cells 28
Mitochondrial DNA: A Barrier To Autologous Cell Therapeutics 29
Induced Pluripotency And The Potential To Save Endangered Species 30
Chapter Summary 33
Key Terms 37
Review Questions 39
Thought Question 40
Suggested Readings 40
2 FUNDAMENTALS OF STEM CELLS 43
Basic in Vitro Cell Culture—A Historical Perspective 43
Stem Cell Culture—Optimal Conditions and Techniques 48
Embryonic Stem Cell Culture 49
Hematopoietic Stem Cell Culture 52
Notch Regulation of HSC Proliferation 52
Other Drivers of HSC Proliferation 53
Adipose]Derived Stem Cell Culture 54
The Study of Embryonic Development 56
Embryonic Development and the Origin of Stem Cells 56
Early Events in Embryogenesis 56
Germ Cell Development 61
Basic Properties of Stem Cells 63
Long]Term Self]Renewal 63
Different Potency Capabilities 63
Totipotency 64
Pluripotency 65
Multipotency 68
Oligopotency 68
Unipotency 69
Types of Stem Cells 70
Embryonic Stem Cells 70
Fetal Stem Cells 70
Amniotic Stem Cells 71
Adult Stem Cells 71
Induced Pluripotency (iPS) Cells 71
Cancer Stem Cells 71
The Potential of Stem Cells in Medicine and Medical Research 71
Therapeutics 71
Tissue Engineering 71
Cell Therapy 73
Cell]Based Drug Screening 75
Chapter Summary 77
Key Terms 80
Review Questions 83
Thought Question 84
Suggested Readings 85
3 EMBRYONIC STEM, FETAL, AND AMNIOTIC STEM CELLS 87
ES Cells 87
Basic Properties 87
Pluripotency 87
Indefinite Replicative Capacity 89
Signaling and Transcriptional Control of ES Cell Replication 90
Examples of ES Cells 92
Mouse ES Cells 92
Rat ES Cells 95
Nonhuman Primate ES Cells 97
Human ES Cells 100
EC Cells 103
Embryonal Germ Cells 105
EG Cell Growth Factor Signaling 105
Comparing Embryonically Derived Cells 106
Fetal Stem Cells 108
Basic Properties 108
Amniotic Fluid Stem Cells 108
Wharton’s Jelly Stem Cells 109
Amniotic Membrane Stem Cells 110
Placental Stem Cells 110
Chapter Summary 112
Key Terms 114
Review Questions 115
Thought Question 116
Suggested Reading 116
4 ADULT STEM CELLS 118
Discovery and Origin of ASCs 118
Basic Properties of ASCs 118
Self]Renewal 119
Multipotency 119
Examples of ASCs 120
Hematopoietic Stem Cells 122
Morphology and Marker Expression 123
Sources 123
Signaling and Multipotency 123
Muscle]Derived Stem Cells 127
Myosatellite Cell Morphology and Marker Expression 129
Sources 129
Signaling, Transcriptional Control, and Multipotency 129
Adipose]Derived Stem Cells 130
Morphology and Marker Expression 130
Sources 131
Signaling and Multipotency 132
Mesenchymal Stem Cells 134
Morphology and Marker Expression 134
Sources 136
Signaling and Multipotency 136
Neural Stem Cells 140
Morphology and Marker Expression 141
Sources and Origins 143
Signaling and Multipotency 146
Endothelial Stem Cells 150
Morphology and Marker Expression 150
Sources and Origins 152
Signaling and Multipotency 153
Chapter Summary 156
Key Terms 158
Review Questions 161
Thought Question 162
Suggested Readings 162
5 NUCLEAR REPROGRAMMING 164
Examples of Nuclear Reprogramming in Nature 166
Cell Fusion 166
Cell Fusion for the Generation of Hybridomas 169
Mechanisms of Cell Fusion 172
Comparison of Cell Fusion Techniques 173
Electrofusion 173
Pegylation 173
Viral Induction 173
Mechanism of Nuclear Reprogramming in Cell Fusion 175
Somatic Cell Nuclear Transfer 176
Method for the Production of SCNT]Derived Cells 177
Somatic Cell Nuclear Transfer for the Creation of Stem Cells 178
Basic Properties of SCNT]Derived Stem Cells 180
Examples of SCNT]Derived Stem Cells 181
A Note Regarding Genomic Abnormalities in SCNT]Derived Clones 182
Telomeric Length 182
DNA Methylation and Epigenetics 183
X]Chromosome Inactivation 185
Induced Pluripotency 185
Breakthrough in the Production of iPS Cells 187
Methods for the Production of iPS Cells 191
Retroviral and Lentiviral Gene Delivery 192
Adenoviral Gene Delivery 195
Sendai Viral Gene Delivery 196
Plasmid]Based Gene Delivery 196
mRNA Delivery 198
MicroRNA Induction 199
Transposon Delivery 200
Direct Protein Delivery 200
Basic Properties of iPS Cells 201
A General Comparison of iPS Cells and Embryonic Stem Cells 203
Examples of Derived iPS Cells 205
Mouse Embryonic Fibroblast]Derived iPS Cells 206
Human Adult Skin]Derived iPS Cells 209
Advantages of ips Cells Over Other Cell Types 210
Origin and Bioethics 211
Patient Specificity 211
Broad Lineage Differentiation Potential 211
“Unlimited” Supply 211
Ease of Generation 212
Chapter Summary 212
Key Terms 214
Review Questions 217
Thought Question 218
Suggested Readings 218
6 CANCER STEM CELLS 220
Background on The Origins of Cancer 220
Discovery and Origin of Cancer Stem Cells 221
Basic Properties of Cancer Stem Cells 225
A Comparison of Cancer Stem Cells and Normal Stem Cells 226
Signaling Pathways Involved in Cancer Stem Cell Transformation 228
Examples of Cancer Stem Cells 229
Breast 229
Central Nervous System 229
Colon 231
Ovary 233
Pancreas 233
Prostate 234
Melanoma 235
Multiple Myeloma 237
Strategies for Treatment Targeting Cancer Stem Cells 238
Melanoma Treatment Targeting Chimeric Antigen Receptor 238
Multiple Myeloma Treatment Targeting CD20 239
Chapter Summary 240
Key Terms 241
Review Questions 243
Thought Question 244
Suggested Readings 244
7 STEM CELLS AS DRUG DISCOVERY PLATFORMS 245
Embryonic Stem Cells and Mouse Models of Gene Function 245
Stem Cell]Based Screening Assays 250
Stem Cells as Lineage Resources for HTS 250
Embryonic Stem Cells as a Resource 251
Adult Stem Cells as a Resource 253
iPS Cells as a Resource 254
Cancer Stem Cell Screens 256
Reprogramming Screens 258
Analysis of Disease Pathways 261
Stem Cells As A Toxicity]Testing Platform 267
Stem Cells as a Resource for Developmental Toxicity Testing 267
Stem Cells as a Source for Post]Natal Environmental Toxicity Testing 268
Cardiotoxicity 269
Hepatotoxicity 272
Chapter Summary 273
Key Terms 274
Review Questions 276
Thought Question 277
Suggested Readings 277
8 THERAPEUTIC APPLICATIONS OF STEM CELLS 279
History of Stem Cells as Therapeutics 279
History of Tissue Engineering 279
Disease]Specific Treatment and Patient Trials 282
Stem Cell]Based Patient Trials: An Overview 282
Cardiomyopathy and Cardiovascular Disease (CV) 284
Neuropathies and Neurodegenerative Diseases 286
Spinal Cord Injury 286
Brain Damage 288
Parkinson’s Disease 292
Autoimmune Disorders 292
Corneal Defects 296
Hematopoietic Disorders 297
Sickle Cell Disease 297
Wiskott–Aldrich Syndrome 299
Cancer 300
Muscular Dystrophy 301
Liver Disorders 303
Veterinary Applications 306
Equine 307
Canine 309
Osteoarthritis (OA) 309
Myocardial Infarction 311
Spinal Cord Injury 313
Stem Cells as an Emerging Industry 314
Seminal Discoveries Driving the Growth of a New Industry 317
Regulation and Reimbursement of Stem Cell Commercialization 320
A Word about Induced Pluripotency and Commercialization 320
Chapter Summary 321
Key Terms 324
Review Questions 326
Thought Question 327
Suggested Readings 328
About the Author 330
Index 331
