The Science of Stem Cells

Introduces all of the essential cell biology and developmental biology background for the study of stem cells  This book gives you all the important information you need to become a stem cell scientist. It covers the characterization of cells, genetic techniques for modifying cells and organisms, tissue culture technology, transplantation immunology, properties of pluripotent and tissue specific stem cells and, in particular, the relevant aspects of mammalian developmental biology. It dispels many misconceptions about stem cells—especially that they can be miracle cells that can cure all ills. The book puts emphasis on stem cell behavior in its biological context and on how to study it. Throughout, the approach is simple, direct, and logical, and evidence is given to support conclusions.  Stem cell biology has huge potential for advancing therapies for many distressing and recalcitrant diseases, and its potential will be realized most quickly when as many people as possible have a good grounding in the science of stem cells. Content focused on the basic science underpinning stem cell biology Covers techniques of studying cell properties and cell lineage in vivo and in vitro Explains the basics of embryonic development and cell differentiation, as well as the essential cell biology processes of signaling, gene expression, and cell division Includes instructor resources such as further reading and figures for downloading Offers an online supplement summarizing current clinical applications of stem cells Written by a prominent leader in the field, The Science of Stem Cells is an ideal course book for advanced undergraduates or graduate students studying stem cell biology, regenerative medicine, tissue engineering, and other topics of science and biology. 

Table of Contents:
Preface xi About the Companion Website xiii 1 What is a Stem Cell? 1 Stem Cell Markers 3 Label‐ Retention 4 The Niche 5 Asymmetric Division and Differentiated Progeny 6 Clonogenicity and Transplantation 6 In Vivo Lineage Labeling 7 Conclusions 9 Further Reading 10 2 Characterizing Cells 13 Histological and Anatomical Methods 13 Histological Sections 13 Fixation 13 Sectioning 14 Staining 14 Electron Microscopy 15 Fluorescence Microscopy 16 Wholemounts 17 Immunostaining 17 In Situ Hybridization 18 Other Methods 19 RNAseq 19 Laser Capture Microdissection 19 Flow Cytometry 20 Dividing Cells 21 The Cell Cycle 21 Studying Cell Turnover 24 Reporters for the Cell Cycle 26 Identification of Very Slow Cell Turnover 26 Classification of Cell Types by Proliferative Behavior 28 Cell Death 28 Further Reading 30 3 Genetic Modification and the Labeling of Cell Lineages 31 Introducing Genes to Cells 31 Transfection and Electroporation 31 Gene Delivery Viruses 33 Controlling Gene Expression 35 Tet System 35 Cre System 35 Inhibiting Gene Activity 37 CRISPR‐Cas9 37 Transgenic Mice 38 Animal Procedures 38 Modification of Embryonic Stem Cells 40 Types of Transgenic Mice 41 Cell Lineage 42 Cell Lineage, Fate Maps, Clonal Analysis 43 Use of CreER for Lineage Analysis 44 Retroviral Barcoding 46 Clonal Analysis in Humans 47 Further Reading 47 4 Tissue Culture, Tissue Engineering and Grafting49 Simple Tissue Culture 51 Media 51 Contamination 53 Growth in Culture 54 Cryopreservation and Banking 55 GMP Cultivation 56 Complex Tissue Culture 56 Induced Differentiation 56 Three Dimensional Cell Culture 57 Artificial Organs and Organoids 59 Grafting 60 The Immune System 61 T Cells 61 The Major Histocompatibility Complex 62 T and B Cell Responses 63 Reactions to a Graft 64 Immunosuppressive Drugs 65 Animal Experiments Involving Grafting 66 Further Reading 67 5 Early Mouse and Human Development 69 Gametogenesis 70 Germ Cells 70 Mitosis and Meiosis 70 Primordial Germ Cells (PGCs) 72 Spermatogenesis 73 Oogenesis 73 Fertilization 76 Early Development 77 Preimplantation Phase 77 Implantation Period – Mouse 80 Implantation Period – Human 82 Ethical and Legal Issues Concerning the Early Human Conceptus 85 Sex Determination 86 X‐Inactivation 87 Imprinting 87 Cloning by Nuclear Transplantation (SCNT) 89 Further Reading 90 6 Pluripotent Stem Cells 93 Mouse Pluripotent Stem Cells 93 Mouse Embryonic Stem Cells 93 Differentiation of Mouse ES cells 95 Mouse iPS Cells 97 Human Pluripotent Stem Cells 101 SCNT‐Derived Embryonic Stem Cells 102 Ethical Issues Concerning Human ES Cells 102 Pluripotent Stem Cells from Postnatal Organisms 103 Applications of Pluripotent Stem Cells 104 Further Reading 105 7 Body Plan Formation 107 Embryological Concepts 107 Developmental Commitment 107 Embryonic Induction 109 Symmetry Breaking 110 Key Molecules Controlling Development 111 Genes Encoding Developmental Commitment 111 Inducing Factors 112 Wnt System 112 FGF System 113 Nodals and BMPs 114 Notch System 114 Hedgehog System 115 Growth Promoting Pathways 115 Retinoic Acid 115 Body Plan Formation 116 General Body Plan 116 Gastrulation 116 Embryo Folding 120 Further Reading 123 8 Organogenesis 125 Nervous System 125 The Brain 126 Regional Specification of the CNS 128 Rostrocaudal 128 Mediolateral 130 Dorsoventral 131 The Eye 131 The Neural Crest 132 Epidermis 134 Hair Follicles 135 Mammary Glands 136 Somitogenesis 137 The Somite Oscillator and Gradient 138 Subdivision of the Somites 139 Myogenesis 140 The Kidney 140 Blood and Blood Vessels 142 Blood 142 Blood Vessels 143 The Heart 145 The Gut 146 Regional Specification of the Endoderm 148 The Intestine 149 The Pancreas 150 The Liver 150 Further Reading 151 9 Cell Differentiation and Growth 155 Organs, Tissues and Cell Types 155 Epithelia 156 Connective Tissues 156 Cell Differentiation 158 Regulation of Gene Activity 158 Lateral Inhibition 161 Asymmetrical Cell Division 162 Neurogenesis and Gliogenesis 164 Neurons and Glia 164 Neurogenesis 166 Gliogenesis 168 Postnatal Cell Division 169 Adult Neurogenesis 169 Neurospheres 171 Skeletal and Cardiac Muscle 171 Skeletal Muscle 171 Development of Skeletal Muscle 172 Muscle Satellite Cells 173 Cardiac Muscle 175 Endodermal Tissues 176 Cell Differentiation in the Pancreas 176 Cell Differentiation in the Intestine 178 Cell Differentiation in the Liver 179 Hepatocytes and Cholangiocytes 180 Liver Growth and Regeneration 181 Transdifferentiation and Direct Reprogramming of Cell Type 183 Differentiation Protocols for Pluripotent Stem Cells 184 Further Reading 185 10 Stem Cells in the Body 189 The Intestinal Epithelium 189 Intestinal Stem Cells 191 In Vitro Culture 193 Clonality of Intestinal Crypts 193 The Epidermis 195 Hair Follicles 197 Cornea and Limbus 199 Mammary Glands 200 Mammary Stem Cells 203 The Hematopoietic System 204 Analysis by Transplantation and in Vitro Culture 204 Hematopoiesis in the Steady State 207 The Hematopoietic Niche 209 Spermatogenesis 211 Further Reading 213 11 Regeneration, Wound Healing and Cancer 217 Planarian Regeneration 217 Neoblasts 218 Amphibian Limb Regeneration 220 The Regeneration Blastema 220 Pattern Formation in Regeneration 222 Mesenchymal Stem Cells 223 Mammalian Wound Healing 225 Soft Tissue Wounds 225 Healing of Bone Fractures 225 Spinal Cord Injuries 227 Regeneration and Repair 228 Cancer 228 Genetic Heterogeneity of Cancer 230 Cancer Stem Cells 233 Further Reading 236 Index 239