Lentropin, a factor present in the vitreous humor of the eye, stimulates lens fiber differentiation from chicken embryo lens epithelial cells in vitro. Lentropin has been partially purified but has not been isolated in sufficient quantity or purity for direct comparison with other growth and differentiation factors. Previous studies have shown that insulin and fetal bovine serum share with lentropin the ability to stimulate lens fiber formation from cultured epithelial cells. In the present study, a number of hormones and growth factors were assayed for lentropin activity. Of those tested, the only substances that had this activity were the insulin-like growth factors (IGFs) somatomedin C (Sm-C/IGF-I) and multiplication-stimulating activity (MSA/IGF-II). Sm-C/IGF-I was approximately 30 times more potent than insulin or MSA/IGF-II in promoting fiber cell formation. A monoclonal antibody to human Sm-C/IGF-I inhibited purified Sm-C/IGF-I, fetal bovine serum, and chicken vitreous humor from stimulating fiber cell differentiation in vitro. This antibody has been shown not to crossreact with insulin and did not block insulin-stimulated lens fiber formation. These findings indicate that lentropin is related to the IGFs and that these factors may play important roles in controlling cell differentiation, in addition to their better-known ability to stimulate cell division.