The recently discovered osteogenic growth peptide (OGP) has been shown to regulate proliferation in fibroblastic and osteoblastic cell lines derived from rats and mice and also alkaline phosphatase activity in the latter was found to be affected. In vivo the OGP enhances bone formation and trabecular bone density. The results of the current study indicate that the OGP is also a potent regulator of marrow stromal cells from man and rabbit, as well as rabbit muscle fibroblasts. The main OGP activity in both marrow systems is a marked stimulation of alkaline phosphatase activity and matrix mineralization. In the rabbit‐derived cell culture this enhancement is accompanied by a reciprocal inhibition of proliferation. On the other hand, the human cells show a concomitant increase of both parameters. The proliferative effect of the OGP is similar to that of growth hormone (GH) and basic fibroblast growth factor (bFGF). The combined activity of the OGP with GH is smaller than that of each of the polypeptides alone. The OGP and bFGF potentiate each other. Of the three polypeptides tested, OGP is the most potent enhancer of alkaline phosphatase activity and mineralization. bFGF has no influence on these characteristics of osteogenic maturation. The OGP maturational activity is unaffected by either GH or bFGF. These data suggest that the marrow stromal cells serve as targets for the OGP that mediate the OGP‐induced increase in osteogenesis. The effect on the human cells implies a role for the OGP in clinical situations where the osteogenic potential of bone marrow is involved.