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Mouse mammary fat pad interacts with mitogens to stimulate epithelial growth in vitro

We have examined the ability of co-cultured mouse mammary fat pad (MFP) to modulate the response of mouse mammary epithelial cells to a number of mitogens in vifro. In a series of 4 experiments, COMMA- 1 D cells were cultured with or without epithelium-free MFP obtained from the abdominal mammary glands of 23-day old virgin, female BALBlc mice. Final cell number was quantified as total DNA after7 days. The main effect of MFP was significant in all experiments, MFP alone inducing a 6.1 to 8.2-fold increase in cell number relative to basal medium (BM) controls (PcO.001). Furthermore, this stimulatory effect of MFPsynergised (PcO.02) with the mitogenic effect of 10% foetal calf serum (FCS). Examination of the response to several mammogenic hormones indicated that insulin (10 @ml) alone did not increase cell number (p>O.O5), and interacted with the effect of MIT (P O.O5), whilst epidermal growth factor (EGF; 25 m 0.5 ug/ml) or indomethacin (5 ug/ml),an inhibitor of prostaglandin synthesis, in the absence or presence of MFP. These findings demonstrate that a diffusible factor(s) from the MFP interacts with several regulators of mammary growth to markedly stimulate epithelial proliferation in vitro. Such a factorappears distinct from anumberofclassical mitogens, and may be involved in the regulation of mammary development in vivo.