A potential therapy for myocardial infarction is to deliver isolated mesenchymal stem cells (MSCs) to the infarcted site. A key issue with this technology is the development of a suitable system for MSC delivery. Our delivery system of interest is a fibrin-based patch used to entrap cells during polymerization. This delivery vehicle lias many advantages; however the mechanical properties and the limited capacity for tailoring cell response may restrict its application. We have developed a PEGylated fibrin patch for MSC transplantation by modifying fibrinogen (Fgn) with the benzotriazole carbonate derivative of PEG to create secondary crosslinking. In this study, the chemical PEGylation of fibrinogen was verified by both amine group quantification and SDS-PAGE. The clotting characteristics and physical properties were compared between the fibrin patch and PEGylated fibrin patch. After seeding with porcine MSCs, the cell viability, morphology, and motility in the novel patch were observed. Phenotypic changes in the embedded MSCs were examined using immunohistochemistry and RT-PCR. The optimal molar ratio (PEG:Fgn = 10:1) was determined for loading MSCs in vitro into the PEGylated fibrin patch. The results suggest that our PEGylated fibrin patch increases MSC viability. Furthermore, the PEGylated fibrin causes phenotypic changes in MSCs consistent with endothelial cells.