Neural Stem Cell Assays
Herausgegeben von Kaur, Navjot; Vemuri, Mohan C.
Neural Stem Cell Assays
Herausgegeben von Kaur, Navjot; Vemuri, Mohan C.
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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
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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 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.
Hinweis: Dieser Artikel kann nur an eine deutsche Lieferadresse ausgeliefert werden.
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.
Hinweis: Dieser Artikel kann nur an eine deutsche Lieferadresse ausgeliefert werden.
Produktdetails
- Produktdetails
- Verlag: Wiley & Sons
- 1. Auflage
- Seitenzahl: 288
- Erscheinungstermin: 9. Februar 2015
- Englisch
- Abmessung: 244mm x 170mm x 23mm
- Gewicht: 640g
- ISBN-13: 9781118308257
- ISBN-10: 1118308255
- Artikelnr.: 39764623
- Verlag: Wiley & Sons
- 1. Auflage
- Seitenzahl: 288
- Erscheinungstermin: 9. Februar 2015
- Englisch
- Abmessung: 244mm x 170mm x 23mm
- Gewicht: 640g
- ISBN-13: 9781118308257
- ISBN-10: 1118308255
- Artikelnr.: 39764623
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.
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
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
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
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