Many of the potentialities in the Tissue Engineering (TE) field, including some of the most promising processing techniques, are somehow hindered by the lack of non-destructive, real-time monitoring tools, capable of reporting on cell molecular status. So far, the majority of techniques used to monitor biological responses in TE approaches rely on destructive and laborious procedures, providing only endpoint information, thus impairing research progression and potential clinical translations. Recently, live-cell imaging with resource to reporter-genes technology appeared as an attractive alternative to monitor biological outcomes. Problems such as photo-bleaching of fluorescent protein or cell damage due to continuous exposition to UV light during cell tracking studies can now be overcome using engineered photo-inducible and photoconvertible fluorescent proteins, such as Dendra2. In this work, we propose the encapsulation of Dendra2-transfected hASCs in microfluidic fibers produced through an oil-free approach compatible with the most common polymers crosslinking strategies. Photo-switching of the fluorescent reporter protein in the cells encapsulated in a 3D environment through confocal microscopy was confirmed. Overall, the proposed strategy will allow not only the assessment of cell retention and migration patterns in fibres but also opens the path to study molecular mechanisms and responses of encapsulated cells in 3D, through the design of specific reporter constructs, acting as molecular switches.
|