Evaluation of CD49e as a distinguishing marker for human articular cartilage derived chondroprogenitors

Background: Cell-based therapy in cartilage repair can benefit from the use of chondroprogenitors; a cell type classified as mesenchymal stem cells, demonstrating reduced hypertrophy. Fibronectin, routinely used to isolate chondroprogenitors, classically binds to alpha 5 beta 1 integrins (CD49e + CD29), of which CD49e is said to be highly expressed in progenitors. The aim of our study was to assess the specificity of CD49e as a distinguishing marker for chondroprogenitors; because studies report low expression in fresh chondrocytes (FCs), but re cent conflicting data has exhibited incremental expression of CD49e in cultured chondrocytes. Methods: FCs were isolated from three human osteoarthritic knee joints and CD49e - cells (sorted by flow cytometry) were cultured in adherent and non-adherent conditions and reassessed for CD49e and CD29 at multiple time points. Colony-forming efficiency (CFE) following fibronectin adhesion assay was calculated for FC, CD49e+ and CD49e - cells. Results: A statistically significant increase in CD49e and CD29 expression was seen in both adherent and non-adherent cultures of CD49e - cells (P < 0.01), as early as 24 h. All groups grew clonally and CFE was similar without any significant difference. CD49e- chondrocytes turned positive when cultured, possibly due to an inherent phenotypic drift, seen after release from cartilage and not because of plastic adherence or chondroprogenitor overgrowth, as non-adherent cultures also showed high expression. Conclusions: As the specificity of CD49e is questionable, there is a pressing need for a specific differentiating marker, to isolate a pure population of chondroprogenitors, as this cell type shows inherent chondrogenesis and reduced hypertrophy, both requisites for cartilage repair. (C) 2020 Elsevier B.V. All rights reserved.