Human Pluripotent Stem Cells

Comprehensive coverage of the entire induced pluripotent stem cell basic work flow

Pluripotent stem cells (PSC) can divide indefinitely, self-renew, and can differentiate to functionally reconstitute almost any cell in the normal developmental pathway, given the right conditions. This comprehensive book, which was developed from a training course, covers all of the PSCs (embryonic, embryonic germ, and embryonic carcinoma) and their functions. It demonstrates the feeder-dependent and feeder-free culture of hESC and hiPSC, which will be referred to in all protocols as PSCs. It also addresses the methods commonly used to determine pluripotency, as defined by self-renewal marker expression and differentiation potential.

Human Pluripotent Stem Cells: A Practical Guide offers in-depth chapter coverage of introduction to stem cell, PSC culture, reprogramming, differentiation, PSC characterization, and more. It also includes four appendixes containing information on reagents, medias, and solutions; common antibodies; consumable and equipment; and logs and forms.

• Includes helpful tips and tricks that are normally omitted from regular research papers
• Features useful images to support the technical aspects and results visually as well as diagrammatic illustrations
• Presents specific sections (ie: reprogramming, differentiation) in a concise and easily digestible manner
• Written by experts with extensive experience in stem cell technologies

Human Pluripotent Stem Cells: A Practical Guide is an ideal text for stem cell researchers, including principal investigators, and others in university and industry settings, and for new graduate students in PSC labs.

1 Introduction 1

1.1 Biosafety 1

1.2 Biosafety Levels 2

1.3 Aseptic Technique 2

1.4 Storage 6

1.5 Contamination 7

1.6 Pluripotent Stem Cells 8

1.7 Procedures 9

References 16

2 Pluripotent Stem Cell Culture 17

2.1 Introduction 17

2.2 Materials 18

2.3 Solutions 19

2.4 Methods 21

References 47

3 Reprogramming 49

3.1 Introduction 49

3.2 Materials 49

3.3 Solutions 54

3.4 Methods 57

References 97

4 Characterization 99

4.1 Introduction 99

4.2 Materials 101

4.3 Solutions 103

4.4 Methods 104

References 128

5 Differentiation 131

5.1 Introduction 131

5.2 Materials 131

5.3 Solutions 133

5.4 Methods 135

References 151

Index 153

Uma Lakshmipathy, Director R&D, Cell Biology, Life Sciences Solutions, Thermo Fisher Scientific, USA

Chad C. MacArthur, Staff Scientist R&D, Cell Biology, Life Sciences Solutions, Thermo Fisher Scientific, USA

Mahalakshmi Sridharan, Scientist R&D, Cell Biology, Life Sciences Solutions, Thermo Fisher Scientific, USA

Rene H. Quintanilla, Scientist R&D, Cell Biology, Life Sciences Solutions, Thermo Fisher Scientific, USA

Comprehensive coverage of the entire induced pluripotent stem cell basic work flow

Pluripotent stem cells (PSC) can divide indefinitely, self-renew, and can differentiate to functionally reconstitute almost any cell in the normal developmental pathway, given the right conditions. This comprehensive book, which was developed from a training course, covers all of the PSCs (embryonic, embryonic germ, and embryonic carcinoma) and their functions. It demonstrates the feeder-dependent and feeder-free culture of hESC and hiPSC, which will be referred to in all protocols as PSCs. It also addresses the methods commonly used to determine pluripotency, as defined by self-renewal marker expression and differentiation potential.

Human Pluripotent Stem Cells: A Practical Guide offers in-depth chapter coverage of introduction to stem cell, PSC culture, reprogramming, differentiation, PSC characterization, and more.

• Includes helpful tips and tricks that are normally omitted from regular research papers
• Features useful images to support the technical aspects and results visually as well as diagrammatic illustrations
• Presents specific sections (i.e., reprogramming, differentiation) in a concise and easily digestible manner
• Written by experts with extensive experience in stem cell technologies

Human Pluripotent Stem Cells: A Practical Guide is an ideal text for stem cell researchers, including principal investigators, and others in university and industry settings, and for new graduate students in PSC labs.