Thesis Jeroen Stein (LUMC): Advanced tools and methods for modeling cardiovascular disease using human pluripotent stem cells

On November 24th 2022 Jeroen Stein successfully defended his thesis, entitled: ‘Advanced tools and methods for modeling cardiovascular disease using human pluripotent stem cells’ at Leiden University. His research was performed under the supervision of Prof. Mummery, Dr. Milena Bellin and Dr. Valeria Orlova.

To advance disease modelling and drug discovery leading to new pharmaceutical options for cardiovascular patients, new in vitro models and quantification methods are needed that improve clinical predictivity. For this purpose several approaches have been explored, ranging from novel co-culture methods to functional read-outs for human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), in order to bring new solutions for disease modeling which suitably reflects aspects of the human heart. Firstly, the current status of the Heart-on-chip (HoC) field and the cutting edge (bio)materials were reviewed, which enable researchers to create more physiologically relevant culture methods for hiPSC-CMs. To this end, a polydimethylsiloxane HoC was used to culture hiPSC-CM, hiPSC-endothelial cells and hiPSC-cardiac fibroblasts combined with a microfluidic flow-channel to mimic physiological blood flow. Subsequently, a silicon wafer-based platform for miniaturized 3D-culture of EHTs designed ultimately for medium- to high-throughput drug screening was fabricated and tested. Moreover, a stand-alone software application was created for automated and robust analysis of in vitro models of striated muscle. A previously published mathematical tool was used and applied to static, live, 2D, 3D, hiPSC-CM, primary CM and skeletal muscle, providing evidence of its versatility. This was successfully applied to study hiPSC-CMs from patients with hypertrophic cardiomyopathy and carrying a heterozygous mutation in the MYBPC3 gene, which encodes a structural protein that is part of the sarcomeres. A broad range of read-outs was used to assess the sarcomeric disarray in these cells, as well as the contractile phenotype in different formats of in vitro cardiac models. Finally, the results and conclusions presented in this thesis have been discussed, and an outlook of future research directions in this exciting field has been provided.

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