Table Of ContentStem Cells in Toxicology and Medicine
Stem Cells in Toxicology and Medicine
Editor
Saura C. Sahu
Center for Food Safety and Applied Nutrition,
US Food and Drug Administration, USA
This edition first published 2017
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Library of Congress Cataloging‐in‐Publication Data
Names: Sahu, Saura C., editor.
Title: Stem cells in toxicology and medicine / [edited by] Saura C. Sahu.
Description: Chichester, UK ; Hoboken, NJ : John Wiley & Sons, 2017. | Includes bibliographical references and index.
Identifiers: LCCN 2016025077 (print) | LCCN 2016039295 (ebook) | ISBN 9781119135418 (cloth) | ISBN 9781119135425 (pdf) |
ISBN 9781119135449 (epub)
Subjects: LCSH: Stem cells–Research. | Toxicology–Research.
Classification: LCC QH588.S83 S74 2016 (print) | LCC QH588.S83 (ebook) | DDC 616.02/774–dc23
LC record available at https://lccn.loc.gov/2016025077
A catalogue record for this book is available from the British Library.
Cover Image: thebackground/Gettyimages
Set in 10/12pt Times by SPi Global, Pondicherry, India
10 9 8 7 6 5 4 3 2 1
Dedication
I lovingly dedicate this book to:
My parents, Gopinath and Ichhamoni, for their gifts of life, love and living example.
My wife, Jharana, for her life‐long friendship, love and support as well as for her patience and
understanding about the long hours spent at home on planning, writing and editing this book.
My children, Megha, Sudhir, and Subir, for their love and care.
Saura C. Sahu
Laurel, Maryland, USA
Contents
List of Contributors xx
Preface xxvi
Acknowledgements xxvii
PART I 1
1 Introduction 3
Saura C. Sahu
References 4
2 Application of Stem Cells and iPS Cells in Toxicology 5
Maria Virginia Caballero, Ramon A. Espinoza‐Lewis, and Manila Candiracci
2.1 Introduction 5
2.2 Significance 6
2.3 Stem Cell (SC) Classification 7
2.4 Stem Cells and Pharmacotoxicological Screenings 8
2.5 Industrial Utilization Showcases Stem Cell Technology as a Research Tool 8
2.6 Multipotent Stem Cells (Adult Stem Cells) Characteristics and Current Uses 9
2.7 Mesenchymal Stem Cells (Adult Stem Cells) 10
2.8 Hematopoietic Stem Cells (Adult Stem Cells) 11
2.9 Cardiotoxicity 12
2.10 Hepatotoxicity 15
2.11 Epigenetic Profile 17
2.12 Use of SC and iPSC in Drug Safety 18
2.12.1 Potential Benefits of Stem Cell Use in Other Areas 18
2.12.2 Methodologies 18
2.12.3 Economic Benefits of Stem Cell Use 19
2.13 Conclusions and Future Applications 19
Acknowledgments 19
References 19
3 Stem Cells: A Potential Source for High Throughput Screening in Toxicology 26
Harish K Handral, Gopu Sriram, and Tong Cao
3.1 Introduction 26
3.2 Stem Cells 27
viii Contents
3.2.1 Embryonic Stem Cells (ESC s) 27
3.2.2 Foetal Stem Cells 29
3.2.3 Adult Stem Cells 29
3.2.4 Adult Stem Cells in Other Tissues 30
3.3 High Throughput Screening (HTS) 31
3.3.1 Current Strategies and Types of High Throughput Screening 32
3.3.2 In Vitro Biochemical Assays 33
3.3.2.1 Fluorescent Based Assays 33
3.3.2.2 Luminescence‐Based Assays 33
3.3.2.3 Colorimetric and Chromogenic Assays 34
3.3.2.4 Mass Spectroscopy (MS) Based Detection Assays 34
3.3.2.5 Chromatography‐Based Assays 34
3.3.2.6 Immobilization and Label‐Free Detection Assays 35
3.3.3 Cell‐Based Assays 35
3.3.3.1 Reporter Gene Assays 36
3.3.3.2 Cell‐Based Label Free Readouts 36
3.4 Need for a Stem Cell Approach in High Throughput Toxicity Studies 37
3.5 Role of Stem Cells in High Throughput Screening for Toxicity Prediction 38
3.5.1 Applications of Stem Cells in Cardiotoxicity HTS 38
3.5.2 Applications of Stem Cells in Hepatotoxicity HTS 39
3.5.3 Applications of Stem Cells in Neurotoxicity HTS 40
3.6 Conclusion 40
Acknowledgement 41
Disclosure Statement 41
Author’s Contribution 41
References 41
4 Human Pluripotent Stem Cells for Toxicological Screening 50
Lili Du and Dunjin Chen
4.1 Introduction 50
4.2 The Biological Characteristics of hPSCs 51
4.2.1 The Biological Characteristics of hESCs 51
4.2.2 The Biological Characteristics of hiPSCs 51
4.3 Screening of Embryotoxic Effects using hPSCs 52
4.3.1 Screening of Embryotoxic Effects using hESCs 52
4.3.2 Screening of Embryotoxic Effects using hiPSCs 54
4.4 The Potential of hPSC‐Derived Neural Lineages in Neurotoxicology 55
4.4.1 T he Challenge of hPSC s‐Derived Neural Lineages in Neurotoxicology
Applications 55
4.4.2 The New Biomarkers in Neurotoxicology using hPSC ‐Derived Neural Lineages 56
4.4.2.1 Gene Expression Regulation 56
4.4.2.2 Epigenetic Markers 57
4.4.2.3 Mitochondrial Function 58
4.4.3 The New Methods in Neurotoxicology using hPSC ‐Derived Neural Lineages 58
4.4.3.1 High‐Throughput Methods 58
4.4.3.2 Three‐Dimensional (3‐D) Culture 59
4.5 The Potential of hPSC ‐Derived Cardiomyocytes in Cardiotoxicity 60
4.5.1 The Challenge of hPSC ‐Derived Cardiomyocytes in Cardiotoxicology Applications 60
Contents ix
4.5.2 The New Biomarkers in Cardiotoxicology using hPSC ‐Derived Cardiomyocytes 60
4.5.2.1 Gene Expression 61
4.5.2.2 Multi‐Electrode Array 61
4.5.3 High‐Throughput Methods 62
4.6 The Potential of hPSC‐Derived Hepatocytes in Hepatotoxicity 62
4.6.1 The Challenge of hPSCs‐Derived Hepatocytes in Hepatotoxicology Application 62
4.6.2 The New Biomarkers in Hepatotoxicology using hPSC ‐Derived Hepatocytes 63
4.6.3 The New Methods in Hepatotoxicology using hPSC ‐Derived Hepatocytes 64
4.6.3.1 iPSC ‐ HH ‐Based Micropatterned Co‐Cultures (iMPCC s) with Murine
Embryonic Fibroblasts 64
4.6.3.2 Suspension Culture of Aggregates of ES Cell‐Derived Hepatocytes 65
4.6.3.3 Long‐Term Exposure to Toxic Drugs 65
4.7 Future Challenges and Perspectives for Embryotoxicity and Developmental Toxicity
Studies using hPSCs 65
Acknowledgments 66
References 67
5 Effects of Culture Conditions on Maturation of Stem Cell‐Derived Cardiomyocytes 71
Deborah K. Hansen, Amy L. Inselman, and Xi Yang
5.1 Introduction 71
5.2 Lengthening Culture Time 75
5.3 Substrate Stiffness 76
5.4 Structured Substrates 78
5.5 Conclusions 82
Disclaimer 82
References 83
6 Human Stem Cell‐Derived Cardiomyocyte In Vitro Models for Cardiotoxicity Screening 85
Tracy Walker, Kate Harris, Evie Maifoshie, and Khuram Chaudhary
6.1 Introduction 85
6.1.1 Cardiotoxicity in Preclinical and Clinical Drug Development 85
6.1.2 Functional Cardiotoxicity 86
6.1.3 Structural Cardiotoxicity 87
6.1.4 Requirement for Improved In Vitro Models to Predict Human Cardiotoxicity 88
6.2 Overview of hPSC‐Derived Cardiomyocytes 88
6.3 Human PSC‐CM Models for Cardiotoxicity Investigations 90
6.3.1 hPSC‐CMs for the Assessment of Electrophysiological Cardiotoxicity 90
6.3.1.1 Patch Clamp Assays 91
6.3.1.2 Voltage Sensitive Dyes (VSDs) 92
6.3.1.3 Optogenetics 93
6.3.1.4 Multielectrode Array (MEA) Assays 94
6.3.1.5 Impedance Assays 96
6.3.1.6 Calcium Imaging Assays 98
6.3.2 hPSC‐CMs for the Assessment of Contractile Cardiotoxicity 98
6.3.2.1 Muscular Thin Films 99
6.3.2.2 Engineered Heart Tissues (EHTs) 99
6.3.2.3 Impedance Assays 101
6.3.2.4 Calcium Imaging Assays 102
x Contents
6.3.3 hPSC‐CMs for the Assessment of Structural Cardiotoxicity 102
6.3.3.1 Mechanisms of Cardiomyocyte Cell Death as Endpoints in Drug Screening 103
6.3.3.2 High Content Analysis 106
6.3.3.3 Impedance Assays 108
6.3.3.4 SeaHorse Flux Analysers 109
6.3.3.5 Complex and 3D Models 110
6.4 Conclusions and Future Direction 112
References 112
7 Disease‐Specific Stem Cell Models for Toxicological Screenings and Drug Development 122
Matthias Jung, Juliane‐Susanne Jung, Jovita Schiller, and Insa S. Schroeder
7.1 Evidence for Stem Cell‐Based Drug Development and Toxicological Screenings
in Psychiatric Diseases, Cardiovascular Diseases and Diabetes 122
7.1.1 Introduction into Stem‐Cell Based Drug Development and Toxicological Screenings 122
7.1.2 Relevance for Psychiatric and Cardiovascular Diseases 123
7.1.3 Advantages of Human Disease‐Specific Stem Cell Models 124
7.1.4 Pluripotent Stem Cell Models 125
7.1.5 Reprogramming of Somatic Cells for Disease‐Specific Stem Cell Models 126
7.1.6 Transdifferentation of Somatic Cells for Disease‐Specific Stem Cell Models 126
7.2 Disease‐Specific Stem Cell Models for Drug Development in Psychiatric Disorders 127
7.2.1 Disease‐Specific Stem Cell Models Mimicking Neurodegenerative Disorder 127
7.2.2 Disease‐Specific Stem Cell Models Mimicking AD 128
7.2.3 Disease‐Specific Stem Cell Models Mimicking Neurodevelopmental Disorders 129
7.2.4 Disease‐Specific Stem Cell Models Mimicking SCZ 131
7.3 Stem Cell Models for Cardiotoxicity and Cardiovascular Disorders 132
7.3.1 Generating Cardiomyocytes In Vitro 132
7.3.2 Generating Microphysiological Systems to Mimic the Human Heart 133
7.3.3 Disease‐Modeling using Microphysiological Cardiac Systems 133
7.4 Stem Cell Models for Toxicological Screenings of EDCs 133
7.4.1 In Vitro Analysis of EDCs in Reproduction and Development 134
7.4.2 In Vitro Analysis and Toxicological Screenings of Drugs 135
References 135
8 Three‐Dimensional Culture Systems and Humanized Liver Models Using Hepatic
Stem Cells for Enhanced Toxicity Assessment 145
Ran‐Ran Zhang, Yun‐Wen Zheng, and Hideki Taniguchi
8.1 Introduction 145
8.2 Hepatic Cell Lines and Primary Human Hepatocytes 146
8.3 Embryonic Stem Cells and Induced Pluripotent Stem‐Cell Derived Hepatocytes 147
8.4 Ex Vivo: Three‐Dimensional and Multiple‐Cell Culture System 148
8.5 In Vivo: Humanized Liver Models 149
8.6 Summary 150
Acknowledgments 150
References 150
9 Utilization of In Vitro Neurotoxicity Models in Pre‐Clinical Toxicity Assessment 155
Karin Staflin, Dinah Misner, and Donna Dambach
9.1 Introduction 155
Contents xi
9.1.1 Limitations of Animal Models and the Utility of In Vitro Assays for Neurotoxicity
Testing 155
9.1.2 How Regulatory Requirements Can Shape the Development of In Vitro Screening
Tools and Efforts 157
9.1.3 In Vitro Assays as Useful Tools for Assessing Neurotoxicity in a Pharmaceutical
Industry Setting 158
9.2 Current Models of Drug‐Related Clinical Neuropathies and Effects on Electrophysiological
Function 159
9.2.1 Neuropathy Assessment 160
9.2.2 Seizure Potential and Electrophysiological Function Assessments 161
9.2.3 Multi Electrode Arrays to Model Electrophysiological Changes Upon
Drug Treatment 161
9.3 Cell Types that Can Potentially Be Used for In Vitro Neurotoxicity Assessment in Drug
Development 162
9.3.1 Primary Cells Harvested from Neuronal Tissues 162
9.3.2 Immortalized Cells and Cell Lines 164
9.3.3 Induced Pluripotent Stem (iPS) Derived Cells 165
9.4 Utility of iPSC Derived Neurons in In Vitro Safety Assessment 167
9.4.1 iPSC Derived Neurons in Electrophysiology 167
9.4.2 iPSC Derived Neurons to Study Neurite Dynamics 167
9.5 Summary of Key Points for Consideration in Neurotoxicity Assay Development 170
9.6 Concluding Remarks 172
References 172
10 A Human Stem Cell Model for Creating Placental Syncytiotrophoblast, the Major
Cellular Barrier that Limits Fetal Exposure to Xenobiotics 179
R. Michael Roberts, Shinichiro Yabe, Ying Yang, and Toshihiko Ezashi
10.1 Introduction 179
10.2 General Features of Placental Structure 180
10.3 The Human Placenta 180
10.4 Human Placental Cells in Toxicology Research 182
10.5 Placental Trophoblast Derived from hESC 183
10.6 Isolation of Syncytial Areas from BAP‐Treated H1 ESC Colonies 185
10.7 Developmental Regulation of Genes Encoding Proteins Potentially Involved in Metabolism
of Xenobiotics 185
10.7.1 Cytochrome P450 Family Members 186
10.7.2 SLC Gene Family Members 188
10.7.3 ATP‐Binding Cassette (ABC) Transporters 189
10.7.4 Metallothionein Family Members 190
10.8 Concluding Remarks 191
Acknowledgments 192
References 192
11 The Effects of Endocrine Disruptors on Mesenchymal Stem Cells 196
Marjorie E. Bateman, Amy L. Strong, John McLachlan, Matthew E. Burow,
and Bruce A. Bunnell
11.1 Mesenchymal Stem Cells 196
11.1.1 Characterization 196
xii Contents
11.1.2 Differentiation 197
11.1.2.1 Adipogenic 197
11.1.2.2 Osteogenic 197
11.1.3 Functions and Activities 197
11.2 Endocrine Disruptors 198
11.2.1 EDC Major Epidemiologic Associations 198
11.2.1.1 EDC Association with Obesity 198
11.2.1.2 EDC Association with Diabetes 199
11.2.2 Challenges with Exposure Study Interpretation in Human Subjects 199
11.2.2.1 Nonmonotonicity of EDC Dose‐Response Curves 199
11.2.2.2 EDC Exposure at Critical Developmental Windows and Association
with Adult Disease 200
11.2.2.3 Effects of Combinations of EDCs 200
11.2.3 Mechanisms of Action of EDCs 201
11.3 Pesticides 201
11.3.1 Organophosphates 201
11.3.1.1 Cell‐Type Specific Effects 201
11.3.1.2 Molecular Effects 203
11.3.2 DDT 205
11.3.2.1 Cell‐Specific Effects 205
11.3.2.2 Molecular Effects 206
11.4 Alkyl Phenols and Derivatives 206
11.4.1 Cell‐Specific Effects 208
11.4.1.1 Effects on Adipocytes and Precursors of Adipocytes 208
11.4.1.2 Effects on Osteoblasts and Precursors of Osteoblasts 209
11.4.2 Molecular Effects 209
11.5 Bisphenol A 211
11.5.1 Cell‐Specific Effects 211
11.5.1.1 Effects on Adipocytes and Precursors of Adipocytes 212
11.5.1.2 Effects on Osteoblasts and Precursors of Osteoblasts 214
11.5.2 Molecular Effects 214
11.6 Polychlorinated Biphenyls 216
11.6.1 Cell‐Specific Effects 217
11.6.1.1 Effects on Adipocytes and Precursors of Adipocytes 217
11.6.1.2 Effects on Osteoblasts and Precursors of Osteoblasts 218
11.6.2 Molecular Effects 220
11.7 Phthalates 221
11.7.1 Cell‐Specific Effects 222
11.7.1.1 Effects on Adipocytes and Precursors of Adipocytes 222
11.7.1.2 Effects on Osteoblasts and Precursors of Osteoblasts 223
11.7.2 Molecular Effects 225
11.8 Areas for Future Research 225
11.9 Conclusions 226
Abbreviations 226
References 228
