A protocol for improving iPSC-cardiomyocyte differentiation reproducibility and maturity by 3D cell printing

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Presented at Australasian Society for Stem Cell Research (ASSCR) Annual Scientific Meeting 2023 by:

Trevor Atkeson*¹, Madhavi Pandya*¹, Alex Volkerling², Drew Neavin¹ and Joseph Powell¹

¹ Garvan Institute of Medical Research, Darlinghurst, Australia
² Inventia Life Science, Alexandria, Australia

*Joint Contribution


Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) are a valuable tool to model patient-specific cardiac development, disease, and drug screening in vitro. Ensuring the accurate and consistent reproduction of in vivo environments is essential. While conventional 2D iPSC-CM models express high cardiac regulatory protein cardiac troponin T (cTnT) levels and demonstrate contractions after 8-14 days of differentiation, 2D models lack tissue-relevant cell-to-cell interactions and cell-extracellular matrix interactions of in vitro cardiac tissue. By better mimicking physiological conditions present in the body, 3D models can offer a more precise tool to predict candidate drug cardiotoxicity or proarrhythmic risks in disease treatment. To create a high throughput drug screening platform, we developed a 3D iPSC-CM differentiation built on Inventia Life Science’s RASTRUM™ Bioprinting Platform. Our 3D workflow improves differentiation to yield iPSC-CMs expressing high levels of cTnT after 14 days of differentiation. These differentiations consistently formed contracting organoid-like 3D cell clusters in a scalable 96-well printed format. Our 3D iPSC-CM protocol is reproducible in different iPSC lines and has a range of customisable variables for further experiment-specific refinement. Our 3D iPSC-CM differentiation is a broadly applicable tool to explore drug cardiotoxicity and model cardiomyocyte development and disease in vitro.


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