1. Cell-Based Assays using Differentiated Human Induced Pluripotent Cells M Z. Cader, M. Graf, M. Burcin, C.F. Mandenius, and J.A. Ross
Part I Differentiation of iPC to Target Cells for Assays
2. Monolayer Generation of Vascular Endothelial Cells from Human Pluripotent Stem Cells Klaus Christensen, Filip Roudnicky, Mark Burcin, and Christoph Patsch
3. Generating Human iPSC-Derived Astrocytes with Chemically Defined Medium for In Vitro Disease Modelling Katharina Janssen, Lamiaa Bahnassawy, Claudia Kiefer, Jürgen Korffmann, Georg C. Terstappen, Viktor Lakics, Miroslav Cik, and Peter Reinhardt
4. Human Induced Pluripotent Stem Cell-Derived Definitive Endoderm Bulk Culture and Hepatic Differentiation Anwar A. Palakkan, Jyoti Nanda, and James A. Ross
5. Production of Cardiomyocytes from Human Pluripotent Stem Cells by Bioreactor Technologies Caroline Halloin, Michelle Coffee, Felix Manstein, and Robert Zweigerdt
6. Improved Protocol of Cardiac Differentiation and Maturation of Pluripotent Stem Cells Imen Jebeniani, Shunli Ding, and Michel Pucéat
7. Human Pluripotent Stem Cell Expansion in Stirred Tank Bioreactors Felix Manstein, Caroline Halloin, and Robert Zweigerdt
8. A Protocol for the One-Step Differentiation of Human Induced Pluripotent Stem Cells into Mature Podocytes Cormac Murphy, Elisabeth Feifel, Paul Jennings, Gerhard Gstraunthaler, and Anja Wilmes
9. Differentiation of hiPS Cells into Definitive Endoderm for High Throughput Screening Teresa Bluhmki, Stefanie Traub, Eva Schruf, James Garnett, Florian Gantner, Daniel Bischoff, and Ralf Heilker
Part II High Throughput Metabolomics and Proteomic Cell-Based Assays
10. LC-MS/MS Based Metabolomics for Cell Cultures Anna Artati, Cornelia Prehn, and Jerzy Adamski
11. Preparation of iPSCs for Targeted Proteomic Analysis Anna Baud, Wendy E. Heywood, Daniel Little, Paul Gissen, and Kevin Mills
12. Measurement of Bile Acids as a Marker of the Functionality of iPSC-Derived Hepatocytes Francesca Mazzacuva, Maëlle Lorvellec, Agostino Cilibrizzi, Kevin Mills, Peter Clayton, and paul Gissen
13. Mass Spectrometry Measurement of Albumin/Alpha Fetoprotein Ratio as an Indicator of iPSC-Derived Hepatocyte Differentiation Francesca Mazzacuva, Maëlle Lorvellec, Agostino Cilibrizzi, Kevin Mills, Wendy Heywood, Peter Clayton, and Paul Gissen
Part III High Content Imaging
14. Seeding Induced Pluripotent Stem Cell-Derived Neurons onto 384-Well Plates Daniel Little, Christin Luft, Oliver Pezzini-Picart, Olukunbi Mosaku, Robin Ketteler, Michael J. Devine, and Paul Gissen
15. High Content Autophagy Analysis in iPSC-Derived Neurons using Immuno-Fluorescence Daniel Little, Christin Luft, Olukunbi Mosaku, Robin Ketteler, Michael J. Devine, and Paul Gissen
16. High Content Analysis of Mitochondrial Function in iPSC-Derived Neurons Daniel Little, Christin Luft, Olukunbi Mosaku, Robin Ketteler, Michael J. Devine, and Paul Gissen
17. Evaluating the Effect of Drug Compounds on Cardiac Spheroids using the Cardiac Cell Outgrowth Assay Jonas Christoppersson, Florian Meier, Henning Kempf, Kristin Schwanke, Michelle Coffee, Mario Beilmann, Robet Zweigerdt, and Carl-Fredrik Mandenius
Part IV Electrophysiology Assays
18. Assay Procedures for Compound Testing of hiPSC-Derived Cardiomyocytes using Multi-Well Microelectrode Arrays Udo Kraushaar and Elke Guenther
19. Multielectrode Array (MEA) Based Detection of Spontaneous Network Activity in Human iPSC-Derived Cortical Neurons Valeria Kizner, Sandra Fischer, and Maximilian Naujock
Part V Miscellaneous Cell-Based Assays
20. Assessing Neuronal Excitability on a Fluorometric Imaging Plate Reader (FLIPR) Following a Defined Electrostimulation Paradigm Valeria Kizner, Sandra Fischer, Benjamin Jähnke, and Maximilian Naujock
21. Fabrication of a Microfluidic Cell Culture Device using Photolithographic and Soft Lithographic Techniques Jonas Christoffersson and Carl-Fredrik Mandenius
22. Using a Microfluidic Device for Culture and Drug Toxicity Testing of 3D Cells Jonas Christoffersson and Carl-Fredrik Mandenius
23. hiPS Cell–Derived Neurons for High-Throughput Screening Stefanie Traub and Ralf Heilker
The volume presents a variety of protocols that will be useful to researchers working in the field of disease modeling, drug discovery, and the cell biology of various tissues. Chapters in this book cover topics such as requirements and preconditions for using human induced pluripotent cell lines in assay development; the culture of iPSCs and iPSC-derived cardiomyocytes by bioreactor technologies; distinctions between hepatocytes, vascular endothelial cells, cardiomyocytes, renal podocytes, and neurons; and high content assays of mitochondrial function and autophagy. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Cell-Based Assay using iPSCs for Drug Development and Testing: Methods and Protocols is a valuable resource for researchers working in this important and developing field of study.