(T1196) AUTOMATION OF FULL WORKFLOW FOR CARDIAC DIFFERENTIATION AND FORMATION OF 3D CARDIAC ORGANOIDS FROM HUMAN iPSC, AND FUNCTIONAL ANALYSIS OF COMPOUND RESPONSES

Abstract: Modeling human tissues using iPSC -derived 3D cardiac organoids is a highly promising technology to facilitate drug development and toxicity assessment. In this study, we developed method to automate the full process for cell differentiation from iPSC and formation and maintenance of 3D cardiac organoids. CellXpress.ai automated cell culture system contains four essential components for automated organoid culture: liquid handler, automated incubator and automated imager, plus scheduling software that that provides automated processing.

Cardiac cells were derived from iPSC which were triggered to differentiation with CellXpress.ai instrument. After 36 hours of differentiation, cells were harvested, then cardiac micro-tissues were created in low attachment U-bottom plates. CellXpress.ai instrument automated cell plating and subsequent media exchanges every 2 days and monitoring by imaging every 24h. 3D microtissues were formed within 48 hours and started to contract spontaneously. After additional maturation for 2 weeks, we evaluated the functional activity of cardiac organoids by recording calcium oscillations after addition of calcium-sensitive dye using a fast kinetic fluorescence (FLIPR Penta). Also, we evaluated the morphology of cardiac organoids and confirmed presence of three cell types: cardiomyocytes, fibroblasts and endothelial cells.

We tested the responses of the microtissues to 20 compounds including modulators of cardiac activity, blockers of ion channels, and panel of known cardiotoxic compounds (CIPA). Tested compounds, including hERG inhibitors, ion channel blockers and other compounds demonstrated changes in the Ca2+oscillation patterns consistent with expected mode of action of tested compounds or toxicity effect. Waveform analysis of patterns provided multiple read-outs including peak frequency, peak amplitude, peak prolongation, irregularity, appearance of secondary peaks and other measurements characterizing modulations of oscillation patterns. Additionally, we characterized the morphology and viability of 3D microtissues by image analysis.
The data presented highlight the utility and biological relevance of using iPSC-derived cell types in 3D microtissues as promising model for screening potential cardiotoxic effects in human cardiac tissues.

Funding Source: Molecular Devices, LLC

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