A Novel 3D Label-Free Monitoring System of hES-Derived Cardiomyocyte Clusters: A Step Forward to In Vitro Cardiotoxicity Testing
Unexpected adverse effects on the cardiovascular system remain a major challenge in the development of novel active pharmaceutical ingredients (API). To overcome the current limitations of animal-based in vitro and in vivo test systems, stem cell derived human cardiomyocyte clusters (hCMC) offer the opportunity for highly predictable pre-clinical testing. The three-dimensional structure of hCMC appears more representative of tissue milieu than traditional monolayer cell culture. However, there is a lack of long-term, real time monitoring systems for tissue-like cardiac material. To address this issue, we have developed a microcavity array (MCA)-based label-free monitoring system that eliminates the need for critical hCMC adhesion and outgrowth steps. In contrast, feasible field potential derived action potential recording is possible immediately after positioning within the microcavity. Moreover, this approach allows extended observation of adverse effects on hCMC. For the first time, we describe herein the monitoring of hCMC over 35 days while preserving the hCMC structure and electrophysiological characteristics. Furthermore, we demonstrated the sensitive detection and quantification of adverse API effects using E4031, doxorubicin, and noradrenaline directly on unaltered 3D cultures. The MCA system provides multi-parameter analysis capabilities incorporating field potential recording, impedance spectroscopy, and optical read-outs on individual clusters giving a comprehensive insight into induced cellular alterations within a complex cardiac culture over days or even weeks.
The joint research project PhenoCor deals with the development of a medical diagnostic device (Clinical Multi Electrode Array, cMEA) for a better and more precise in-vitro stratifization, model-based therapy planning and clinical safety pharmacology for patients suffering from a genetically caused heart disease.
Our partners of the University of Leipzig and the University Clinic in Dresden are focusing on the assay and experimental design Sciospec`s role is to develop and build a massiv-multichannel system combining electro-physiological potential recording (EPR), electrical impedance spectroscopy (EIS), electrical impedance tomography (EIT) and complex stimulation within the same system. The system allows for fully simultaneous acquisition of impedance or potential signals on up to 512 electrodes in combination with up to 4 independent complex stimulation injections. This opens up new possibilities for monitoring highly dynamic and spatially distributed electro-physiological phenomena and allows in-depth insights into complex cell interactions. An application-specific sensor adapter brings together the measurement system and the newly designed high density sensor array with an innovative contacting scheme.
PhenoCor is supported by the Free State of Saxony and the European Union (SMWA/EFRE).
Project number: 100387683