The objective of this study was to investigate whether meniscus-derived decellularized matrix (DCM) has the capacity to induce differentiation of synovial fluid-derived mesenchymal stem cells (SF-MSCs) towards a meniscus fibrochondrocyte (MFC) phenotype. The potential roles of transforming growth factor beta-3 (TGF-β3) and insulin-like growth factor 1 (IGF-1) in the differentiation of SF-MSCs towards an MFC phenotype were also investigated. SF-MSCs were isolated via plastic adherence cell culture from the synovial fluid of five donors (5 male, average age 34 years). Porous DCM was generated by homogenizing and freeze-drying fresh normal human cadaveric meniscus tissue. SF-MSCs were seeded and cultured on the DCM scaffold in a defined serum-free media (SFM) supplemented with or without the combination of TGF-β3 and IGF-1. Cell pellets of SF-MSCs were cultured in SFM with either TGF-β3 or IGF-1 or their combination as controls. The duration of culture was 3 weeks for both experimental configurations. We assessed newly-formed tissues by biochemical assays, scanning electron microscopy (SEM), immunofluorescence and quantitative real-time PCR (qPCR). The combination of TGF-β3 and IGF-1 induced production of the cartilaginous matrix in DCM and upregulated the expression of aggrecan, collagens I and II. Moreover, the SF-MSCs exhibited a round morphology in the DCM scaffolds in the presence of the growth factors. In pellets, combined TGF-β3 and IGF-1 synergistically enhanced cartilaginous matrix production. In contrast to bone marrow mesenchymal stem cells (BM-MSCs), the differentiated SF-MSCs showed little evidence of the expression of the hypertrophic differentiation marker, collagen X. In conclusion, meniscus-derived DCM appears to require exogenous growth factor supplementation to direct differentiation of SF-MSCs. STATEMENT OF SIGNIFICANCE: Meniscus tears are the most common injury of the knee joint. These tears pose a major risk factor for the early development of knee osteoarthritis. Unfortunately, the majority of these tears occur in the inner region of the meniscus and lacks blood supply with no reparative or regenerative capacity. The goal of this study was to determine if the native extracellular matrix (ECM) of human meniscus has the capacity to differentiate human knee synovial fluid resident mesenchymal stem cells (SF-MSCs) towards a meniscus phenotype as a potential strategy to repair avascular meniscal tears. Our findings show that the human meniscus-derived ECM without supplementation with growth factors (TGF-β3 and IGF-1) cannot differentiate SF-MSCs towards a meniscus phenotype. The use of meniscus-derived scaffolds as a material to stimulate endogenous repair of meniscus tears via differentiation of SF-MSCs may require supplementation with TGF-β3 and IGF-1.Keywords: Decellularized matrix; Meniscus; Stem cells; Synovial fluid; Tissue engineering.