Main description:

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.

Contents:

Preface...
Table of Contents...
Contributing Authors...

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, Jurgen 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 Puceat

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, Maelle 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, Maelle 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 Jahnke, 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