PET/EVOH nonwoven fabrics support in vitro expansion of mesenchymal stem cells with high differentiation potential

Cell expansion under three-dimensional (3D) condition has been shown to better preserve cellular properties by mimicking the native microenvironment, thereby creating a more physiologically relevant culture system. This study investigated the capacity of a three-dimensional nonwoven polyethylene terephthalate (PET)/ethylene vinyl alcohol (EVOH) scaffold as a 3D substrate for large-scale expansion of mesenchymal stem cells (MSC). A seven-fold increase in cell number was observed after 14 days of cultivation, and cells were well distributed with an efficient infiltration within scaffolds based on the hematoxylin and eosin (H&E) staining. A high cell survival and pertaining of metabolic activity were demonstrated by live/dead and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) stainings, respectively. The preservation of MSC properties was confirmed by their differentiation potential toward osteogenic, adipogenic, and chondrogenic lineages. Both <i>in situ</i> differentiation of cell-expanded scaffolds and the subsequent differentiation after cell retrieving from scaffolds revealed the successful responsiveness of expanded MSC to lineage-specific stimuli. These findings suggest the potential of this PET/EVOH scaffold as the 3D culture substrate enabling efficient MSC proliferation while maintaining key functional properties.