Camila Vesga Castro – Doktoratu ondoko ikertzailea Biodonostia OIIan
Biodonostia OIIko-Ekitaldi Aretoan
For years, standard 2D cellular cultures and animal models have been used to understand muscle physiopathology and to develop novel treatments for muscle diseases. Despite the many discoveries regarding those models, a comparison with native human muscle suggests that major challenges remain in order to obtain a fully representative or trustworthy model. Moreover, the characterization of these in vitro tissues has focused on analyzing myogenic markers, determining the fusion index, and the presence of striation patterns, among others. Whereas this information is helpful for understanding muscle development, functional outcomes such as contractility and calcium handling provide further physiological information about the degree of tissue maturation and functionality. In this context, we first performed a systematic review of the diverse platforms used for the evaluation of contractile force (CF) in in vitro skeletal muscle models. From this analysis, it was evident the necessity of CF normalization for reliable comparison among studies and the identification of key parameters such as tetanic-to-twitch ratio that provides unique information about muscle CF performance. Second, we describe the development of 2D and 3D in vitro skeletal muscle platforms that support muscle organization and the evaluation of functional outcomes. A 2D in vitro culture platform was developed to generate single, isolated myotubes based on a micro groove pattern (µGrooves). Myotubes within µGrooves responded to electrical stimulation and exhibited spontaneous contractions. This platform prompted improved myotubes contractile behavior compared to standard controls without any mechanical cue. Finally, for 3D, two different hydrogel protocols (fibrin and collagen) were tested in order to optimize the formation of bioartificial muscles (BAMs) from immortalized human cells.