For the first time, we demonstrate here the ability of human relaxin to block cell division. During the induction of differentiation of 3T3-L1 fibroblasts to adipocytes, the cells typically undergo two rounds of cell division followed by accumulation of lipid droplets and expression of insulin-stimulated glucose transport as the cells attain the adipocyte phenotype. Human relaxin added during induction had no effect on the development of the adipocyte phenotype or insulin-stimulated glucose transport. However, it blocked cell division at a half-maximal concentration of 1.25 nM, well within physiological range. This could be reversed by the addition of antibodies specific for human relaxin. Thus relaxin joins a select number of hormones with growth inhibitory properties such as transforming growth factor-beta (TGF beta) and mammastatin. Potentially, this is an important but until now unidentified function of relaxin. Unlike other inhibitory polypeptides, like TGF beta, relaxin does not prevent differentiation but rather uncouples it from cell division.