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Polyurethanes (PUs) are able to degrade into harmless molecules upon implantation and have received a significant level of attention as a biomaterial. PUs based on polycaprolactone (PCL) and amino acid derivatives are examples of these polymers. As a potential biomaterial, their biocompatibility coupled with biodegradability attracted attention in tissue engineering applications. In this study, a biodegradable thermoplastic, poly(ester-urethane) based on L-lysine diisocyanate (LDI) and PCL was synthesized. The resulting polymer was fully characterized with nuclear magnetic resonance spectroscopy (NMR), FTIR-ATR, gel permeation chromatography (GPC), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and mechanical test. The surface properties were studied by goniometer and biocompatibility was evaluated using L929 mouse fibroblast cell line. The FTIR-ATR, and NMR results together provide a proof for the synthesis of the polyurethane polymer. According to thermal analysis results, melting temperature and weight loss were found to be around 48°C and between 300°C-450°C, respectively. Cell culture studies showed that the cells attached and proliferated well and had proper morphology on PU films.