Synovial fluid mesenchymal stromal cells (SF-MSC) ignite interests for cartilage tissue engineering. However, inducing chondrogenic differentiation with growth factors can lead to hypertrophic or osteogenic phenotypes. Therefore, assessing new markers occurring during this differentiation is primordial. Our objective was to characterize SF-MSC VS chondrocytes using a classic chondrogenic index completed by an epitranscriptomic mapping along their chondrogenic differentiation to identify new markers. SF-MSC and chondrocytes obtained from osteoarthritic patients were characterized using histology, immunohistochemistry and RT-qPCR completed by rRNAs epitranscriptomic mapping. Cells were characterized during monolayer expansion, pre-treatment (MSC only) and chondrogenic differentiation in collagen sponges treated with TGF-β1 or not (ITS) for 7, 14, 21, 28 and 56 days. A pool of P0 chondrocytes served as reference for analyses. SF-MSC expressed chondrogenic genes and formed a cartilage like ECM under TGF-β1 in collagen sponges. TGF-β1 treated SF-MSC showed lower rRNA methylation rates than non-treated ones at D7. rRNA methylation rate increased after D7 for TGF-β1 treated cells reaching similar levels as control at D56. Five differentially methylated sites were identified between chondrocytes and SF-MSC. This epitranscriptomic mapping allowed to show new chondrogenic markers and targets to control chondrogenic differentiation. This work was supported by the Société Francaise de Rhumatologie