Doctoral Student KTH Royal Institute of Technology KTH Royal Institute of Technology, Stockholms Lan, Sweden
Abstract: The rare disease group of Muscular Dystrophy is incurable genetic diseases where the dystrophin gene has been compromised which can range from mild to severe forms. Muscular Dystrophy mainly manifests itself during early childhood and causes progressive muscle deterioration which leads to a high risk of death due to eventual respiratory or cardiac failure. Disease modelling and drug development currently rely on animal models which fail to accurately recapitulate the human disease mechanism. To address the need for a more accurate patient-derived cell model, a human induced pluripotent stem cell (hiPSC)-approach has been investigated with the goal of generating cardiomyocyte organoids with retained disease phenotype. To better represent in vivo environments, the novel FN-silk scaffold comprising a recombinant spider silk protein functionalized with a cell adhesion motif from fibronectin was used to form 3D culturing systems. Cell-integration and cultivation compatibility of both healthy wild-type hiPSC and disease carrying isogenic hiPSC lines has been reproducibly confirmed in various 3D FN-silk formats such as fibers and free-floating networks, complemented with extracellular matrix proteins. hiPSCs integrated in FN-silk retained pluripotency and proliferative growth which have been confirmed with comparative live/dead viability assays and pluripotency marker staining using immunocytochemistry. Furthermore, hiPSC-derived cardiomyocyte progenitors in 3D FN-silk were generated with a cardiomyocyte differentiation kit. Cardiac markers and beating frequency were used for evaluation of differentiation parameters such as maturity. Although further optimization of cultivation and differentiation parameters is needed, this proof of concept study represents a significant first step for hiPSC-derived rare disease modelling while exploring alternative 3D hiPSC-culturing systems.
