Understanding the role of the pancreatic extracellular matrix (ECM) in supporting islet survival and function drives the pursuit to create biomaterials that imitate and restore the pancreatic ECM microenvironment. To create an ECM mimic holding bioinductive cues for β-cells, self-assembled peptide amphiphiles (PAs) inscribed with four selected ECM-derived cell adhesive ligands are synthesized. After 7 days, compared to control groups cultured on biologically inert substrates, MIN6 β-cells cultured on PAs functionalized with YIGSR and RGDS cell adhesive ligands exhibit elevated insulin secretion in responses to glucose and also form β-cell clusters. These findings suggest that the self-assembled PA nanomatrix may be utilized to improve pancreatic islet transplantation for treating type 1 diabetes. As an extracellular matrix (ECM) mimetic scaffold capable of replicating the native β-cell microenvironment, self-assembled peptide amphiphiles (PAs) inscribed with cell adhesive ligands found within the islet basement membrane proteins are studied. Interestingly, MIN6 cells cultured on substrates that have YIGSR and RGDS cell adhesive sequences, respectively exhibit significantly improved insulin secretion responses to glucose and tend to form β-cell clusters. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.