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Extracellular matrix (ECM) including fibronectin (FN) and elastin plays a pivotal role in providing a microenvironment to support tissue regeneration in stem cell therapy.To develop a novel biomimetic ECM for stem cell differentiation,we engineered FN type Ⅲ 9 and 10 domains fused to elastin-like polypeptides (FN-ELPs).The recombinant FN-ELP fusion protein was expressed in Escherichia coli and purified by inverse transition cycling.Human mesenchymal stem cells (hMSCs) cultured on plates coated with FN-ELP had significantly greater adhesion activity and proliferation than cells grown on non-coated plates.FN-ELP induced the osteogenic differentiation by elevating alkaline phosphatase (ALP) and mineralization activity of hMSCs.Furthermore,the osteogenic marker gene expressions of ALP,collagen type Ⅰ (Col Ⅰ),osteopontin (OPN),and transcriptional coactivator with a PDZ-binding motif (TAZ) were increased in hMSCs cultured on plates coated with FN-ELR We reported a novel biomimetic ECM with potential for bone regeneration that promotes the osteogenic differentiation of hMSCs.