Although it is well known that the metabolism of chondrocytes can be altered by the application of mechanical strain, it is unclear whether chondrocytes from the superficial and deep zones of cartilage respond in a similar manner. In this study, chondrocytes from the uppermost 15-20% (superficial cells) and the remaining tissue (deep cells) from bovine articular cartilage were isolated separately and cultured in agarose constructs. Cell deformation on application of a 15% static compressive strain was identical for both subpopulations after 24 and 72 hours in culture. The constructs were incubated under static and dynamic (0.3, 1, and 3 Hz) strains of 15% amplitude. Glycosaminoglycan synthesis by deep cells was unaffected by static strain or 3 Hz dynamic strain, whereas 0.3 Hz produced a significant reduction and 1 Hz induced a highly significant 50% stimulation of glycosaminoglycan synthesis (p < 0.001). Superficial cells exhibited a general inhibition of glycosaminoglycan synthesis. By contrast, proliferation of superficial cells was stimulated by dynamic strain whereas deep cells were not influenced. It has been suggested previously that mechanotransduction-induced controls of glycosaminoglycan synthesis and proliferation in chondrocytes embedded in agarose are uncoupled. Data presented in this study demonstrate that the two processes do, in fact, occur in different subpopulations of chondrocytes within the full-depth cell isolate.