Three-dimensional mesenchymal stromal cells spheroids generated by RGD-induced self-assembly technique for tissue...

Recently, generation of 3D spheroids from mesenchymal stromal cells (MSCs) for tissue engineering has been proposed, in order to recapitulate in vivo-like conditions, namely mild hypoxia and proper cell-cell interactions. The aim of the current study was to develop a novel simple approach to generate multicellular spheroids directly from monolayer cell culture of mesenchymal stromal cells. For this purpose, cell self-aggregation induced by the cyclo-RGDfK(TPP) peptide (5-100 µM) has been proposed. The cells were shown to form spheroids after adding the peptide to culture medium within 72 h. The spheroids were stained with vital dye Calcein AM, and cell viability was confirmed by confocal laser scanning microscopy. The spheroids were able to reverse into single-cell culture after peptide removal, which provided cell invasion into the collagen hydrogel. The spheroid-based cultivation approach could be promising for tissue engineering.