Monitoring the maturation of the sarcomere network: a super-resolution microscopy-based approach

• DOI: 10.1007/s00018-022-04196-3
• Publication Date: 2022-02-23T18:03:25Z
• Authors: Anna Skorska; Lisa Johann; Oleksandra Chaban...; Praveen Vasudevan; Sophie Kussauer; Maximilian Hillem...; Markus Wolfien; Anika Jonitz-Heincke; Olaf Wolkenhauer; Rainer Bader; Hermann Lang; Robert David; Heiko Lemcke
• Keywords: Regenerative Medicine

The in vitro generation of human cardiomyocytes derived from induced pluripotent stem cells (iPSC) is great importance for cardiac disease modeling, drug-testing applications and regenerative medicine. Despite the development various cultivation strategies, a sufficiently high degree maturation still decisive limiting factor successful application these cells. process includes, among others, proper formation sarcomere structures, mediating contraction cardiomyocytes. To precisely monitor contractile machinery, we have established an imaging-based strategy that allows quantitative evaluation important parameters, defining quality network. iPSC-derived were subjected to different culture conditions improve formation, including prolonged time micro patterned surfaces. Fluorescent images α-actinin acquired using super-resolution microscopy. Subsequently, determined cell morphology, density, filament alignment, z-Disc thickness length Cells adult neonatal heart tissue served as control. Our image analysis revealed profound effect on content orientation when cultured structured, line-shaped Similarly, had beneficial structural maturation, leading more adult-like phenotype. Automatic filaments by machine learning validated our data. Moreover, successfully transferred this approach skeletal muscle cells, showing improved over differentiation periods. Overall, image-based workflow can be used straight-forward tool quantitatively estimate As such, it support establishment novel protocols enhance maturity.