The deliverable includes the optimization of human induced Pluripotent Stem Cells (hiPSCs) culture and their differentiation in Cardiomyocytes (CMs), the development of the 3D cell system to assess cardiotoxicity (and the testing of chemicals in this system). The first step concerned the assessment of the cell differentiation protocol of hiPSCs in CMs. The second step included the co-culture of Human Coronary Artery Endothelial Cells (HCAECs) with hiPSC- CMs for 14 days. Mitochondrial activity, ROS, LDH, and molecular biology markers were tested, and samples were provided to WP5 for omics. In collaboration with WP2, 3, 5, and 6, Doxorubicin (DOX) was selected as the gold standard of cardiotoxicity and tested to establish a dose-response curve in the standard 2D culture of hiPSC-CMs for a preliminary evaluation of the dose and timing to be used for WP6 analysis. The same dose-response experiment was performed in the 3D co-culture of hiPSC-CMs and HCAECs in Gelatin-Methacryloyl (GelMA). Mitochondrial activity, ROS, LDH, Cardiac Troponin, p53, and ET-1 were tested. For data analysis, samples were provided to WP5 for omics and data to WP6.
ALTERNATIVE - D4.3 Testing results in 3D culture
2023
Abstract
The deliverable includes the optimization of human induced Pluripotent Stem Cells (hiPSCs) culture and their differentiation in Cardiomyocytes (CMs), the development of the 3D cell system to assess cardiotoxicity (and the testing of chemicals in this system). The first step concerned the assessment of the cell differentiation protocol of hiPSCs in CMs. The second step included the co-culture of Human Coronary Artery Endothelial Cells (HCAECs) with hiPSC- CMs for 14 days. Mitochondrial activity, ROS, LDH, and molecular biology markers were tested, and samples were provided to WP5 for omics. In collaboration with WP2, 3, 5, and 6, Doxorubicin (DOX) was selected as the gold standard of cardiotoxicity and tested to establish a dose-response curve in the standard 2D culture of hiPSC-CMs for a preliminary evaluation of the dose and timing to be used for WP6 analysis. The same dose-response experiment was performed in the 3D co-culture of hiPSC-CMs and HCAECs in Gelatin-Methacryloyl (GelMA). Mitochondrial activity, ROS, LDH, Cardiac Troponin, p53, and ET-1 were tested. For data analysis, samples were provided to WP5 for omics and data to WP6.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
