ArticleJournal of translational medicine2026
Modulating mesenchymal stromal cells chondrogenesis through TGF-β1 pre-conditioning during monolayer expansion in pellet culture and in a 3D mechanical load model.
Article in Journal of translational medicine, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
backgroundBone marrow-derived mesenchymal stem cells (MSCs) represent a promising source for cell-based regenerative cartilage therapies. However, in vitro chondrogenic differentiation of MSCs varies widely between donors, posing a significant challenge to the clinical translation of these therapies. Here, we explored the potential of pre-conditioning MSCs with TGF-β1 during monolayer expansion in order to enhance chondrogenic differentiation in poorly responsive MSCs and reduce donor-to-donor variability.
methodsMSCs from six donors were expanded in monolayer using standard methods. For the final two days of expansion, half of the cells were pre-conditioned with 10 ng/mL TGF-β1, whereas the others were kept in standard expansion medium. MSCs were then detached for 28 days of chondrogenic differentiation in standard pellet culture in chondrogenic medium or in a 3D mechanically induced chondrogenesis model in polyurethane-fibrin scaffolds. After 28 days, samples were collected for gene expression analysis, sulphated glycosaminoglycan (sGAG) assay, DNA quantification and histological evaluations (Safranin-O/Fast green staining).
resultsTwo days of TGF-β1 pre-conditioning during MSC expansion increased COL2A1 expression after 28 days of pellet culture (P = 0.0217), but also elevated COL10A1 (P = 0.0293), leaving the COL2A1/COL10A1 ratio unchanged (P = 0.8522). There was a trend for an increased COL2A1/COL1A1 ratio and an increased ACAN expression (P = 0.0837). Pre-conditioning had no effect on sGAGs retained in the pellets or released into the medium over the 28 days but slightly enhanced the Safranin-O staining on histological sections. When chondrogenesis was mechanically induced without exogenous TGF-β1, pre-conditioning had no effect on COL2A1, COL10A1, COL1A1, or ACAN gene expression. Similarly, no differences were observed in sGAG content (retained or released) or in Safranin-O staining.
conclusionsTGF-β1 pre-conditioning enhanced some chondrogenic markers in static pellet culture, but these effects were small and did not translate to the more clinically relevant model of mechanically induced chondrogenesis. Furthermore, TGF-β1 pre-conditioning was associated with increased COL10A1 expression, indicating a concurrent promotion of hypertrophic differentiation.
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