The use of mechanical stimulation during tissue engineering of bone is under much investigation. Stimulation of cells in constructs can lead to improved osteogenic differentiation and extracellular matrix production of mesenchymal stem cells. Low magnitude, high frequency (LMHF) vibration has been used in vivo to improve bone quality in patients with low bone mass and shows promise for improving the healing of large defects. As the mechanisms behind this stimulation are unknown, in vitro research has begun to investigate the response of different cell types found in bone to LMHF vibration. This research studies the osteogenic differentiation of a human mesenchymal progenitor cell line following this type of stimulation under a variety of conditions. Cell attachment to a 3D scaffold is demonstrated, before the use of LMHF vibration in 2D and later in 3D using mechanically different scaffolds. A possible signalling mechanism is also investigated. The research shows positive effects on both early cell differentiation and matrix mineralisation. Variability in the response of cells to loading regimens is found, suggesting that the responses to this type of loading are very sensitive to environmental conditions.