Stem Cells

Rob Burgess, PhD is Vice President of Global Business Development for RayBiotech, Inc. He is also an Assistant Professor of Molecular and Cell Biology at the University of Texas at Dallas within the School of Natural Sciences. He has co-founded several successful start-up companies (Lexicon Genetics, Inc. and Medical Nanotechnologies); restructured and streamlined research, development and manufacturing efforts at Zyvex Corporation; managed over 35 R&D and manufacturing scientists and engineers at Zyvex Corp; authored several book chapters and one textbook on nanomedicine, and published several papers in major peer-reviewed journals such as Science and Nature. He is an expert in the fields of vertebrae functional genomics, stem cells and nanotechnology, with a specific emphasis on stem cell gene targeting and knockout technologies.

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