Cell Behavior on Peptide-Immobilized Substrate with Cell Aggregation Inducing Property

Abstract

Cell aggregates have been applied in various fields such as regenerative medicine and drug toxicity testing. We have shown that H-(Lys-Pro)12-OH (KP24), a repeating sequence of lysine (Lys) and proline (Pro), induces uniformly sized cell aggregates simply by its presence in cell suspension. In this study, we considered that this peptide could be applied to a three-dimensional culture substrate that can induce uniform cell aggregates by immobilizing it on the substrate. Therefore, mouse fibroblasts (L929) were seeded on KP24-immobilized glass substrates and cell behavior was observed. We also seeded human-derived cells, namely, human mesenchymal stem cells (hMSC), on KP24-immobilized substrates and characterized their cell assemblies. Furthermore, KP24 was chemically immobilized on the substrate surface, which allowed us to trace the mechanism of KP24–cell interaction. As a mechanism analysis of the cell aggregation ability of KP24, we investigated whether KP24 interacts with the cell surface without being incorporated into the cell.