Adult bone-marrow-derived mesenchymal stroma cells (BMSC) seem to be a potential cell source for tissue engineering of the ligament. The objective of this work was to study the time-related changes in mRNA expression and protein levels of collagen types I and III and of tenascin-C in BMSC under co-culture with fibroblasts or under a uniaxial cyclic condition. Rat BMSC harvested from the femur and tibial bone marrow were co-cultured with ligament fibroblasts or stimulated by cyclic 10% uniaxial stretching at 1 Hz. Image analysis showed significant cell loss in stretched BMSC, particularly in the directions close to the stretching direction. However, these BMSC displayed an equivalent growth rate to that of non-stretched cells. Real-time reverse transcription/polymerase chain reaction revealed that the mRNA expression of collagen types I and III and of tenascin-C by BMSC was significantly up-regulated by co-culture and cyclic stretching. Radioimmunoassay results confirmed the effects of these stimulations, showing increases in the level of these proteins. Thus, BMSC might be useful as a cell source for the tissue engineering of ligament.