Bioactive factors, such as TGF ß and BMP-2, as well as mechanical factors i.e. compressive loading and hydraulic pressure, have been shown to induce and/or modulate chondrogenesis of bone marrow derived mesenchymal stromal cells (BMSCs). Since these factors are intracellularly transduced through different mechanisms, it is hypothesized that TGF ß, BMP-2 and hydraulic pressure may act synergistically on chondrogenic differentiation of BMSCs. Aggregates of bovine BMSC were cultured in the presence of 10 ng/ml TGF ß1, 50 ng/ml BMP-2 or both. Half of the samples were loaded for 4 hours per day with 0.5-3 MPa cyclic hydraulic pressure at 1 Hz. After 14 days of culture/loading, gene expression of chondrogenic genes was assessed. DNA as well as glycosaminoglycan (GAG) content of the pellets were analysed. Neither pressure nor BMP-2 had an influence on GAG/DNA content. However, cells responded to the presence of TGF ß1 with an up-regulation of chondrogenic genes and GAG/DNA of the aggregates increased compared to controls demonstrating the cells ability to respond to external stimuli. The used concentrations of BMP-2 and parameters for pressure were neither able to induce nor modulate chondrogenesis of bovine BMSCs and thus no synergistic effects were observed. © 2009 - IOS Press and the authors. All rights reserved.