2014 CDI European User Group Meeting
Advances in Human iPSC-derived Cell Models
for Phenotypic Screening & Toxicity Studies
This free multisession conference featured speakers from pharma, academia, and biotechnology presenting data showing the utility of human iPSC-derived iCell® technology in their research.
Bringing Biologically Relevant Cells to Life – Application of iPSC Technology in Lead Discovery and Safety Assessment
Alexander Kvist, PhD
Induced Pluripotent Stem Cell-derived Cardiac Myocytes for the Generation and Electro-architectural Characterisation of Mature Cardiac Tissue In Vitro.
Cesare Terracciano, PhD
Imperial College London
Abstract: Induced pluripotent stem cell-derived cardiac myocytes (IPSC-CM) are a promising tool for cardiac disease modelling, pharmacological and toxicological screening and regenerative strategies. However, their immature, foetal phenotype reduces their applicability and limits their use in cardiac research. The general goal of our research is to define ideal conditions and monitoring strategies for the maturation of IPSC-CM in vitro, in order to obtain a suitable platform to be employed in the screening of the electrophysiological properties of human disease.
Use of Stem Cell-derived Sensory Neurons for Pain Drug Discovery at the Neusentis Research Unit
Peter Stacey, PhD
Functional Evaluation of Human iPSC-derived Dopaminergic Neurons Growing on Micro-electrode Arrays – A Novel In Vitro Screening Platform for Parkinson’s-relevant Drugs
Benjamin Bader, PhD
Abstract: Dopaminergic (DA) neuronal cultures have been widely used for in vitro cell-based assays modeling the pathology of Parkinson’s disease with the goal to screen for novel drug candidates effective to rescue or prevent the induced damage of DA neurons. One of these models is the MPP+ induced oxidative stress model, which specifically damages DA neurons. Our research aims to transfer our current model using primary murine DA neurons to a model using human iPSC-derived DA neurons. We evaluate the spontaneous activity development and the responsiveness to MPP+ of these human DA neurons using multi-well micro electrode arrays analyzed by multi-parametric data analysis and discuss a proof of principle for the use as a screening platform.
Production of Functional Islet Cells from hiPSC: A Step Towards Personalised Nutrition
Marine Kraus, PhD
Nestlé Institute of Health Sciences Lausanne
Abstract: A renewable source of human b-cells would greatly benefit basic research and cell therapy. The in vitro production of pancreatic endoderm (PE) from human stem cells and induced pluripotent stem cells (hiPSC) has been successfully demonstrated (ViaCyte). However, in vitro full maturation of b-cell is rather poor. To achieve this, we are transferring the differentiated pancreatic endoderm (PE) derived from human iPS cells in vivo with the goal of producing human pancreatic models that resemble the human pancreas not only in regards of glucose response and insulin production but also for the whole metabolic behavior of pancreatic cells. Over time, encapsulated iPSC-derived glucose responsive cells not only regulate glucose levels but protect mice against STZ-induced hyperglycemia. All together, these findings can pave the way for future “disease in a dish” humanized and personalized models for cell therapy and screening purposes.
Human and Rat Neuronal Microtissues for Toxicity Testing
David Fluri, PhD
Abstract: Currently in vivo animal models are primarily employed to assess for neurotoxic effects of chemicals and potential candidate compounds in drug development. In vitro testing is limited to HTS compatible low complexity 2D cultures of primary brain cells or cell lines and to testing on more sophisticated low throughput explant cultures. We report here the generation of two different scaffold-free 3D microtissue models derived from human and rat cell origins. Rat neuronal microtissues display incorporation of the three major types of brain cells: neurons, astrocytes and oligodendrocytes and are stable over extended time periods (>4 weeks). Myelination is demonstrated by electron microscopy analysis as well as positive immunohistochemical staining of myelin basic protein (MBP), a major constituent of the axon-enwrapping myelin sheet. The human microtissue model consists of iPS-derived astrocytes in co-culture with iPS-derived neurons. Tissue constructs exhibit stable three-dimensional architecture and display positive staining for the neuronal marker β-III-tubulin and the astrocyte marker GFAP.
Future strategies to assess neurotoxicity in vitro will mainly depend on improved predictivity of the models but also on the capacity to increase throughput of such systems.
Characterization of Single and Co-cultured Human iPSC-derived Cardiomyocytes in Scaffold-free 3D Culture
Inselspital, Universitätsspital Bern
Abstract: Rodent cardiomyocytes in 2D cell culture have been the standard in vitro model in basic cardiovascular research and toxicology so far, but there is a need for better models. Both cardiomyocytes and cardiac fibroblasts play important roles in cardiac remodeling by altering their size, morphology, signaling, and function. Hence, a 3D cell culture model using human induced pluripotent stem cell (iPSC)-derived cardiomyocytes and cardiac fibroblasts was developed and characterized.
Engineered Heart Tissue – A Versatile In Vitro Model
Arne Hansen, PhD
UKE Hamburg Eppendorf
Multi-electrode Array Recording and Calcium Ratiometry in hiPSC-derived Cardiomyocytes
Georg Rast, PhD
Technology Panel Discussion:
Standardizing Stem Cell Derived Cardiomyocyte Use: Pharma and Regulatory Consortia Aim to Define Best Practices, Relevant Endpoints, and Higher Throughput Uses
Yama Abassi, PhD, ACEA Biosciences
James Ross, PhD, Axion Biosciences
Godfrey Smith, PhD, Clyde Biosciences
Abstract: The age of limiting cardiac electrophysiology studies to single endpoints, late stage safety studies, and/or only employing animal tissue cells is coming to an end. Advances with in vitro recapitulation of human cardiomyocyte physiology and predictive higher throughput assays are supplementing, and in some cases supplanting, traditional myopic studies employing single ion channels and/or non-human tissues. Such a paradigm shift requires a concerted standardization and validation of use and endpoints across platforms. Large-scale efforts in Japan and the United States (US) are underway to determine best practices for using and implementing stem cell derived cardiomyocytes across higher throughput platforms in discovery, safety and regulatory experiments. TF-2 in Japan and the Comprehensive in vitro Proarrhythmia Assay (CiPA) consortia in the US are a combination of pharma, regulatory, cell provider, contract research organization (CRO), and academic entities united by a common theme of extracting relevant and robust data from stem cell cardiomyocytes by developing industry-wide best practices. This panel discussion will present an update on above consortia progress followed by presentations on each of three higher throughput platforms for interrogating cardiomyocyte biology, impedance measurements, microelectrode arrays (MEA), and voltage-sensitive dyes. The talks will be followed by a question and answer session.
Kinetic Neurite Outgrowth Study of Human iPSC-derived Neurons in Mono- and Co-culture Measured with Long Term, Live-cell Imaging
S. L. Alcantara, J. Brown, P. Garay, L. McGillicuddy, T. O’Callaghan, T. Dale, D. Appledorn, O. McManus, V. Groppi & D.J. Trezise
Essen BioScience, Welwyn Garden City, UK & Ann Arbor, Michigan, 48108 USA
CDI’s workshop, Advances in Human iPSC-derived Cell Models for Phenotypic Screening & Toxicity Studies, brought together researchers in drug development and toxicology for this unique, one-day event. Selected presentations from this workshop are available for download, below.
The Potential for Induced Pluripotent Stem Cell Technology in Drug Discovery and Development
Devyn Smith, PhD, Executive Director – Chief Operating Officer, Neusentis Pfizer
The pharmaceutical industry has undergone a tremendous shift over the last few years due to years of underperformance by R&D. The focus of the industry now is to better predict efficacy and safety of compounds early, enabling quick-kills. We will discuss where iPS cells can enable pharmaceutical R&D and give specific examples across multiple disease areas.
Comparison of Electrophysiological Data from Human Induced Pluripotent Stem Cell Derived
Cardiomyocytes (hiPSC-CMs) to Current Preclinical Cardiac Safety Assays
hiPSC-CMs studied on multielectrode arrays were pharmacologically validated using a set of reference compounds (E4031, nifedipine, verapamil, cisapride, terfenadine, flecainide, mexiletine and quinidine) and results compared to other preclinical cardiac safety assays (patch clamping, rabbit ventricular wedge assay, rodent and non-rodent in vivo models) used to assess cardiovascular electrophysiological risk. Our results show that hiPSC-CMs demonstrate relevant pharmacological responses and show good correlation to other preclinical cardiac safety assays. Based on this data, hiPSC-CMs could provide an excellent alternative in vitro model to screen for drug induced cardiovascular electrophysiological risk and therefore reduce the number of animals currently used in in vitro studies.
Pharmacological Characterization of Ligand- and Voltage-gated Ion Channels Expressed in iCell® Neurons
Daniel Ursu, PhD, Senior Research Scientist, Eli Lilly
The use of inducible pluripotent stem cells to generate neuronal cells is revolutionizing our ability to access and study human neurons and/or glial cells from living subjects and in particular from patients with various neuro-psychiatric diseases. The presentation will focus on the pharmacological characterization of an already established cellular model of human iPSC-derived neurons (iCell® Neurons) and a comparison with data obtained from rodent primary neurons. Functional studies, performed mainly by using calcium flux assays, will be presented for different classes of ligand-gated ion channels: glutamate (AMPA and NMDA), GABA-A, and α7- acetylcholine nicotinic receptors. Data obtained with similar techniques will also be presented for voltage-gated sodium and calcium channels.
Advancements in the Use of iPS Cell-derived Model Systems for In Vitro Disease Modeling and Phenotypic Screening
Induced pluripotent stem cell (iPSC) technology enables the creation of biologically relevant in vitro cell models for discovery of novel therapeutic targets and phenotypic screening approaches to drug development. Here, we present case studies demonstrating the use of iCell Cardiomyocytes and iCell Neurons for disease modeling, target validation, and drug screening. Specifically, these case studies include screening assay development for modulators of cardiac hypertrophy; high-throughput assay application for neuronal synaptic function; and assay design for compound screening and target validation for the prevention of β-amyloid induced toxicity in neurons. These examples provide evidence of the rapid implementation of human iPSC-derived cell models into drug discovery campaigns.
Pharmacology of iPSC-derived Cardiomyocytes in Comparison to Adult and Neonatal Preparations
Sian Harding, PhD, Imperial College London
We are using human embryonic and iPSC-derived cardiomyocytes (hESC-CM/iPSC-CM) to study adrenergic responses, and will discuss their similarities and differences with other commonly used preparations. β1AR and β2AR responses are clearly present in hESC-CM, showing advantages over animal preparations for modeling of adult human cardiomyocyte responses, and iPSC-CM including iCell reproduced this well. Gs/Gi and adenylyl cyclase components of this system are also functional. Hypertrophic responses through the αAR are less robust, with a systematic switch in α1AR isoforms during differentiation in both hESC-CM and iPSC-CM, as well as subtle differences in intracellular signaling between the two cell types.
iPSC & iPSC-derived cells – Value and Application in Drug Discovery
Utilizing biologically and disease-relevant human cells in physiological in vitro cell models plays an important role in the drug discovery process. However, there are well-known limitations in terms of the supply of human primary cells for in vitro studies, limited proliferative capacity, large batch variations and challenges maintaining the phenotype. Human induced pluripotent stem cells (hiPSC) open up possibilities for better predictive efficacy and safety of compounds early in the drug discovery process. Recent progress in the production of hiPSCs and in technologies that can be applied to these cells enable the generation of biologically relevant human cells in large scale. Here, we will discuss the value and application of hiPSCs in drug discovery, present some early validation results using hiPSC derived cardiomyocytes in a proliferation study for cardiac regeneration, and the potential to use the cells to explore cardiac toxicity, and cardiovascular risks.