Neural Stem Cell Assays

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.

Auteur

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.

Contenu
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 ...