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Neural stem cells offer a valuable model system for delineating the cellular and developmental processes in normal and diseased states of the central nervous system. In particular, neural stem cells have huge potential in regenerative medicine, owing to their expansion capability in culture and the ability to differentiate into multiple sub-neural lineages. Neural Stem Cell Assays provides a detailed and comprehensive review of the basic methods for neural stem cell cultures. Including an overview of progress in the field over the past decade, Neural Stem Cell Assays is a one-stop reference…mehr
Neural stem cells offer a valuable model system for delineating the cellular and developmental processes in normal and diseased states of the central nervous system. In particular, neural stem cells have huge potential in regenerative medicine, owing to their expansion capability in culture and the ability to differentiate into multiple sub-neural lineages. Neural Stem Cell Assays provides a detailed and comprehensive review of the basic methods for neural stem cell cultures. Including an overview of progress in the field over the past decade, Neural Stem Cell Assays is a one-stop reference for consistent methods and reliable tools that span the entire assay work flow, from isolation or generation of neural stem cells to characterization, manipulation and final application of neural stem cells in disease paradigms such as Parkinson's disease, multiple sclerosis and amyotrophic lateral sclerosis. An excellent source of information for academic, pharmaceutical and biotechnology researchers who are new to the neural stem cell field, Neural Stem Cell Assays is an invaluable to experienced users who wish to integrate newly developed tools and technologies into their workflow. The book also covers important course material for students at the undergraduate and graduate level who are learning the basics of neural stem cell cultures, and differentiation to sub-neural lineages.
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Navjot Kaur, Ph.D, is Staff Scientist in the Life Technologies division of the Invitrogen Corporation. His research experience covers a wide range of cell biology, protein biochemistry and molecular biology concepts, with special expertise in cell culture and cryopreservation media development, cell-based assays, primary cells, and stable cell lines. Mohan C. Vemuri, Ph.D., MBA is director of stem cell research and development in the Life Technologies division of Invitrogen & Applied Biosystems. The author of designs of peer-reviewed articles in the field, Dr. Vemuri is also a co-editor of three books on stem cells and stem cell cultures.
Inhaltsangabe
Contributors xv Preface xix 1 Neural Differentiation of Pluripotent Stem Cells 1 Mohan C. Vemuri 1.1 Introduction 1 1.2 NSC Derivation from Rosette Formation of Embryoid Bodies 1 1.3 Rosette Free NSC in a Monolayer Culture 2 1.4 Pathways Involved in Neural Tube and Neural Crest Lineages 2 1.5 Differentiation and Gene Expression in Human Brain Development 3 1.6 Conclusions 6 References 6 2 The History and Design of Assays for the Identification and Characterization of Neural Stem Cells 9 Sharon A. Louis and Brent A. Reynolds 2.1 Introduction 9 2.2 Materials 12 2.3 Methods 14 2.4 Notes 21 Acknowledgments 23 References 23 3 Culture of Human Neural Stem Cells 25 Soojung Shin, Yiping Yan and Mohan C Vemuri 3.1 Introduction 25 3.2 Materials 25 3.3 Methods 30 3.4 Notes 33 3.5 Expected Results 33 References 34 4 GMP4-Compatible Production and Expansion of Human Neural Stem Cells 35 David J. Brick, Alexander E. Stover, Hubert E. Nethercott, Maria G. Banuelos and Philip H. Schwartz 4.1 Introduction 35 4.2 Materials 36 4.3 Methods 40 4.4 Notes 50 Acknowledgments 52 References 52 5 Primary Rat Neural Cultures 55 Navjot Kaur 5.1 Introduction 55 5.2 Materials 55 5.3 Methods 57 5.4 Notes 59 References 59 6 Cryopreservation of Human Neural Stem and Progenitor Cells 61 Profico Daniela Celeste, Sgaravizzi Giada, Projetti Pensi Massimo, Muzi Gianmarco, Ricciolini Claudia, Gelati Maurizio and Vescovi Angelo Luigi 6.1 Introduction 61 6.2 Materials 62 6.3 Methods 62 6.4 Notes 65 References 65 7 Cryopreservation and Recovery of Primary Rat Neural Cells 67 Rhonda Newman and Navjot Kaur 7.1 Introduction 67 7.2 Materials 67 7.3 Methods 68 7.4 Notes 70 References 70 8 Efficient Induction and Scale up Generation of Neural Stem Cells from Human Pluripotent Stem Cells 71 Yiping Yan, Soojung Shin and Mohan C. Vemuri 8.1 Introduction 71 8.2 Materials 71 8.3 Methods 73 8.4 Notes 78 References 79 9 Directed Differentiation of Human NSC/NPC into Dopaminergic Neurons 81 Andrzej Swistowski and Xianmin Zeng 9.1 Introduction 81 9.2 Materials 82 9.3 Methods 83 9.4 Notes 88 References 88 10 In vitro Differentiation of Pluripotent Stem Cells towards either Forebrain GABAergic or Midbrain Dopaminergic Neurons 91 Brigham J. Hartley, Bradley Watmuff, Cameron P.J. Hunt, John M. Haynes and Colin W. Pouton 10.1 Introduction 91 10.2 Materials 91 10.3 Methods 94 10.4 Notes 97 10.5 Typical Results 97 References 98 11 Generation of Astrocytes from Human Pluripotent Stem Cells using a Defined System 101 Atossa Shaltouki and Xianmin Zeng 11.1 Introduction 101 11.2 Materials and Methods 101 11.3 Notes 108 References 108 12 Directed Differentiation of Human PSC into Oligodendrocytes 111 Anu Hyysalo and Susanna Narkilahti 12.1 Introduction 111 12.2 Materials 111 12.3 Methods 113 12.4 Notes 116 Acknowledgments 117 References 117 13 Directed Differentiation towards Human Neural Retinal Cells 119 Deepak A. Lamba 13.1 Introduction 119 13.2 Materials 120 13.3 Methods 121 References 126 14 Directed Differentiation of Photoreceptors and Retinal Pigment Epithelium from Adult Mouse Retinal Stem Cells 129 Brian G. Ballios, Valeria Marigo and Derek van der Kooy 14.1 Introduction 129 14.2 Materials 130 14.3 Methods 131 14.4 Notes 134 Acknowledgments 134 References 134 15 Induction of Schwann Cells from Rat Bone Marrow Mesenchymal Stem Cells 137 Shohei Wakao, Masaaki Kitada and Mari Dezawa 15.1 Introduction 137 15.2 Materials 138 15.3 Method 141 15.4 Notes 149 References 151 16 Neural Cell Viability Assays 155 Bonnie J. Hanson and Michael K. Hancock 16.1 Introduction 155 16.2 Materials 157 16.3 Methods 158 16.4 Notes 165 References 165 17 Genetic Modification of Pluripotent Stem Cell Derived Human Neural Progenitor Cells 167 Jamie M. Chilton and Steven L. Stice 17.1 Introduction 167 17.2 Materials 168 17.3 Methods 170 17.4 Notes 176 Acknowledgments 178 References 178 18 Electrophysiology of Neurons 179 Christine M. Emnett and Steven Mennerick 18.1 Introduction 179 18.2 Materials 180 18.3 Methods 181 18.4 Notes 185 References 187 19 Induction and Transplantation of Neural Stem Cells from Human Pluripotent Stem Cells in Experimental Models (Rat) of Parkinson's disease 189 Makoto Motono, Asuka Morizane and Jun Takahashi 19.1 Introduction 189 19.2 Materials 189 19.3 Methods 191 19.4 Notes 194 Acknowledgments 194 References 194 20 Transplantation of Human Neural Stem Cells and Progenitors in Animal Models of Disease 197 Joseph F. Bonner and Itzhak Fischer 20.1 Introduction 197 20.2 General Principles of Effective Transplantation 198 20.3 Different Methods of Transplantation 204 20.4 Materials and Method 205 20.5 Conclusions 211 20.6 Notes 212 Acknowledgments 212 References 212 21 Morphometric Assays for Neural Differentiation and Synaptic Development of Human iPSC-Derived NSC 215 Barbara Calabrese, Leif Dehmelt and Shelley Halpain 21.1 Introduction 215 21.2 Materials 216 21.3 Method 218 21.4 Notes 222 Acknowledgments 223 References 223 22 BacMam Transduction of Neural Stem Cells 225 Rene Quintanilla and Uma Lakshmipathy 22.1 Introduction 225 22.2 Materials 225 22.3 Methods 227 22.4 Notes 228 Acknowledgments 229 References 229 23 The Proximity Ligation Assay: A High Throughput Technique for Protein Analysis in Neuroscience 231 Michela Zaltieri, PierFranco Spano, Cristina Missale and Arianna Bellucci 23.1 Introduction 231 23.2 Materials 232 23.3 Methods 232 23.4 Notes 238 23.5 Troubleshooting 238 Acknowledgments 240 References 240 24 Quantification of Micro-RNA Expression in Human Neural Stem Cells using TaqMan Assays 241 Xavier Nissan and Alexandra Benchoua 24.1 Introduction 241 24.2 Materials 242 24.3 Methods 244 24.4 Notes 248 References 249 25 Genetic Reporter Cell Lines: Tools for Stem Cell Biology and Drug Discovery 251 Cameron P.J. Hunt, Bradley Watmuff, Brigham J. Hartley, Colin W. Pouton and John M. Haynes 25.1 Pluripotent Stem Cell Reporter Lines 251 25.2 Choice of PSC Genetic Reporter Systems 251 25.3 Reporter Systems 252 25.4 Multiple Reporter Lines 254 25.5 Use of Genetic Reporter Escs to Study Neural Differentiation 255 References 260 Index 261
