hiPS-derived differentiated cells for modelling human development and disease

The discovery of hiPSCs made them an appropriate and compelling candidates for disease modelling and personalized cell therapies. The employment of hiPSCs could be a very important tool to better understand pathologies poorly characterized as the ultrarare Cardiospondylocarpofacial syndrome (CSCF). CSCF is a multifactorial syndrome that causes many dysfunctions at the expense of multiple organs. We reported the reprogramming and characterization of a new hiPSC line registered as UNIBS17-A, derived from a patient’s fibroblasts, who was diagnosed for cardiospondylofacial syndrome derived from a de novo mutation c.737-7A>G. We used the Sendai virus as vector for introducing OSKM factors. We obtained a new stable hiPSC line with pluripotent features able to differentiate in the three germs layers, that could be use in disease modelling. hiPSCs are a potential constant source of human cells, in particular hard-to-reach tissue cells such as cardiomyocytes. One of the major limitation in the use of hiPSC for disease modelling is their maturation status. We investigated the expression of the inhibitory isoform of the troponin complex. Our study demonstrates that hi-CMs never fully acquire an adult phenotype, indeed the adult isoform of the inhibitory troponin (cTnI) starts to appear, but the fetal isoform of the inhibitory troponin (ssTnI) is never switch-off both in molecular and protein level, even when cells are cultured for long periods (90 days). Alongside the analysis of hi-CMs maturation, another factor of interest in research is the development of new easily applicable and non-invasive methods to investigate the functionality of hi-CMs in vitro. Here we reported an imaging method that, by exploiting an algorithm (previously described), allow us to monitor the kinetic and dynamic properties of cells over time, without perturbing them. This analysis allows us to monitor frequency, contractility, kinetic energy and force of contraction developed by the hi-CMs kept in culture for 90 days. The data obtained show that the trend over time of the parameter considered in our study could be associated whit the inability of hi-CMs to reach a complete maturation stage. Finally, another hiPSCs field of great interest is to use 3D self-aggregates structure to improve the phenotype of hiPS-derived cells. We derived a 3D model of both spheroids and organoids to better simulate an environment comparable to the organism one. Furthermore, we used a bioreactor (LiveBox) to create a dynamic culture that, leading a continuous flow of nutrient, mimic the natural condition. Our results carried out that mimicking the organs’ characteristics leads cells to acquire a phenotype more similar of those in the human body. Further improvement will be needed, but likely, this system should be a good model on which performing hepatotoxicity test, with the aim of reducing the use of animal models in research.