A new hydrogel for the production of cell spheroids using a quick-gelation method

Peptide-based hydrogels have extraordinary applications in biomedicine and offer an excellent mimic for the 3D microenvironment of the extracellular matrix. A common strategy to produce hydrogels is by the self-assembly of low-molecular weight peptides containing cell adhesion motifs found in proteins such as fibronectin, laminin and osteopontin. In this work we propose the formation of nanostructured scaffolds using the sequence DGRL4DGW, a self-assembling peptide containing motifs from fibronectin (DGR) and epitopes with the capacity to bind with fibronectin (DGW), as well as the application of the resulting hydrogel in supporting the growth of living cells by the self-assembly of a hydrogel made quick gelation in the presence of HFIP (hexafluoro-2-isopropanol) and characterized by classical biophysical techniques such as small angle scattering X-ray (SAXS), Small angle neutron scattering (SANS), Circular dichroism (CD), Atomic force microscopy (AFM), Electron microscopy (EM) and fluorescence. Cytocompatibility assays were performed with HeLa and NIH-3T3 cells using the Calcein-AM technique and the morphology was accessed by the use of confocal microscopy. The resulting hydrogel was not only thermostable with remarkable biophysical qualities, but also induced the differentiation of living cells into spheroids, which can be used for many applications in the biotechnology and tissue engineering field1.

de Mello LR, Carrascosa V, Rebelato E, Juliano MA, Hamley IW, Castelletto V, Vassiliades SV, Alves WA, Nakaie CR, da Silva ER. Nanostructure formation and cell spheroid morphogenesis of a peptide supramolecular hydrogel. Langmuir. 2022 Mar 11;38(11):3434-45.

Acknowledgements: This work was supported by Comissão de Aperfeiçoamento de Ensino Superior (CAPES), Fundação de Apoio a Pesquisa do Estado de São Paulo (FAPESP) and by Universidade Federal de São Paulo – Escola Paulista de Medicina. (UNIFESP)