RECEPTOR-BINDING AND GROWTH-PROMOTING ACTIVITY OF INSULIN-LIKE GROWTH FACTOR-I IN A BOVINE MAMMARY EPITHELIAL-CELL LINE (MAC-T3)

Insulin-like growth factor-I (IGF-I) has been known to be mitogenic to a variety of cell types, although a growth-regulatory role for IGF-I on bovine mammary epithelial cells has not been fully investigated. In the present study, we examined the receptor binding of IGF-I and its effect on growth in a bovine mammary epithelial cell line (MAC-T3). Specific receptors for IGF-I were detected on cultured bovine mammary epithelial cells. Competitive binding revealed that half-maximal inhibition of I-125-labelled IGF-I binding by IGF-I was approximately 3-mu-g/l. Dissociation rate constant of the IGF-I receptor was 3.10+/-0.06 nmol/l (S.E.M.) with a receptor site concentration of 366+/-8 fmol/mg protein for the average of three experiments. IGF-I exerted a positive mitogenic effect on MAC-T3 cells according to both direct DNA assay and thymidine incorporation assay. Moreover, the mitogenic effect of IGF-I on MAC-T3 cells was enhanced by the addition of fetal calf serum in the culture media. The present results suggest that the bovine mammary epithelial cell line (MAC-T3) provides a useful model system with which to study the biological actions of insulin-like growth factors on the bovine mammary secretory tissue in vitro.