A Novel 3D Label-Free Monitoring System of hES-Derived Cardiomyocyte Clusters: A Step Forward to In Vitro Cardiotoxicity Testing
| Autor: | Andrea A. Robitzki, Silvia Vinz, Daniella Steel, Heinz-Georg Jahnke, Stephan Fleischer, Randy Kurz, Peter Sartipy, Kerstin Dahlenborg, Diana Seidel |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Drugs and Devices
Drug Research and Development Time Factors Predictive Toxicology lcsh:Medicine System stability 030204 cardiovascular system & hematology Toxicology Cardiovascular Bioinformatics Cardiotoxins Cardiovascular Pharmacology 03 medical and health sciences 0302 clinical medicine In vivo Molecular Cell Biology Humans Myocytes Cardiac lcsh:Science Biology Embryonic Stem Cells Cell Aggregation 030304 developmental biology Label free 0303 health sciences Cardiotoxicity Multidisciplinary Staining and Labeling Chemistry Stem Cells lcsh:R Signal Processing Computer-Assisted Monitoring system Cell aggregation In vitro Electrophysiological Phenomena 3. Good health Electrophysiology Medicine lcsh:Q Cellular Types Research Article Developmental Biology Biomedical engineering |
| Zdroj: | PLoS ONE; Vol 8 PLoS ONE, Vol 8, Iss 7, p e68971 (2013) PLoS ONE |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0068971 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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