Growth regulation of mouse mammary tumor cells in collagen gel cultures by diffusible factors produced by normal mammary gland epithelium and stromal fibroblasts.

We have reported previously that both normal mammary epithelium and stroma stimulate the growth of mouse mammary tumors in vivo. We have devised a method to investigate the role of diffusible factors in these growth interactions in vitro. Because an appropriate matrix, a particular cell shape, or multicellular organization may be required for the production of factors and/or for the response to such factors, the method assesses the expansion of boluses of cells in collagen gel matrix. Under these culture conditions, paracrine effects were detected which were not observed when cells were growing in monolayer on plastic. The growth of mammary tumor lines 66, 410.4, and D2A1 was stimulated by both mammary epithelial cells and fibroblasts prepared from midpregnant mouse mammary glands. In contrast, these mammary tumor lines were inhibited by coculturing with normal mammary cells in monolayer on plastic. Stimulation in collagen and inhibition in monolayer cultures were both dose dependent; increasing numbers of regulator mammary cells increased the effects on tumor growth. Additional tumor cells did not stimulate growth of target tumor cell boluses in collagen gel cultures. This attempt to model the in situ situation may be a more appropriate model for the detection and characterization of relevant diffusible growth regulatory factors than monolayers on plastic and/or colonies growing in agar.