Apoptosis of primary fibroblasts was observed in vivo during wound healing in skin and is expected to occur in other organs as well; however, the environmental signal for induction of apoptosis in fibroblasts and the putative influence of cell–matrix interactions on the regulation of apoptosis remain to be identified. Here we provide evidence for the role of fibrillar collagen in this process, and demonstrate that normal human primary fibroblasts embedded in contractile collagen gels undergo apoptosis as shown by the appearance of cytoplasmatic histone-associated DNA fragments starting at day 1 of culture with a peak around days 2–4. This induction of apoptosis in primary fibroblasts seems to be specific for contractile collagen gels, because apoptosis of primary fibroblasts was neither observed in cells grown on culture dishes or on plastic dishes coated with collagen, nor observed in cells seeded in either anchored collagen gels or contractile fibrin gels. We therefore conclude that a distinct environment such as a contractile collagen matrix determines the susceptibility of normal primary fibroblasts to apoptosis.