Pectin hydrogels crosslinked via peptide nanofibers for designing cell-instructive dynamic microenvironment

As has been reported many times before, the two-dimensional (2D) cell culture techniques used today are far from modeling native tissue environments. Therefore, tremendous amounts of effort were devoted to developing three-dimensional (3D) cell cultures with high tissue resemblance. Whereas, these techniques suffer from elaborate preparation processes, batch-to-batch variations, unnatural components, chemical modifications, side products, static culture conditions, or complex reactor systems. To overcome these limitations, we report an undocumented one-step strategy to create a tissue-like 3D cell culture method by mimicking the extracellular matrix (ECM) microenvironment with rapid, non-covalent cross-linking of biopolymer-peptide complex and recently designed non-static cell culturing modules. In the current method, we prepared a very facile and tailorable ECM-like network by using easily attainable building blocks without the need for chemical modifications and possible undesirable/noncontrollable responses resulting from these unnatural modifications. Cells encapsulated in this new biopolymer mesh were located in the swimming culture module to mimic not only the microenvironment but also the non-static physical environment of the ECM. The feasibility of this method was analyzed on a bio-regeneration model; SaOS-2 cells cultured in the current 3D system induced improved osteogenic regeneration. The ECM resemblance of the method was also exhibited by histological sections of the cells incubated in the recent gel formulation. Furthermore, different cell types derived from various tissues could be cultured in our recent ECM model, which could be very practicable for personalized test models for future applications as a replacement for animal studies.