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The Science of Stem Cells

ISBN: 978-1-119-23515-6
264 pages
January 2018, Wiley-Blackwell
The Science of Stem Cells (1119235154) cover image

Description

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. 

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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 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 Grafting 49

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

References 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

References 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

 

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Author Information

JONATHAN M. W. SLACK is a developmental biologist and author of five books and over 200 scientific papers. He is an emeritus professor at the University of Bath, UK, as well as the University of Minnesota, USA, where he was Director of the Stem Cell Institute from 2007–2013.

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