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Growth of epithelium from a preneoplastic mammary outgrowth in response to mammary adipose tissue

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We investigated the effects of conditioned media derived from mouse mammary fat pads on the proliferation of CL-S1 cells, an epithelial cell line originally isolated from a preneoplastic mammary outgrowth line. Cell proliferation in vitro in serum-free defined medium was compared to that in this medium conditioned using intact mammary fat pad pieces or isolated fat pad adipocytes. Culture medium was conditioned by incubating the conditioning material in defined culture medium for 24 h at 37°C. Conditioned medium induced CL-S1 proliferation as much as 10- to 20-fold above the minimal levels of growth in control cultures after 13 d of culture. The growth-stimulatory factor(s) had an apparent molecular weight of greater than 10 kDa. This growth-stimulatory activity was both heat and trypsin stable. Because the role of adipose tissue is to store and release lipids, we next tested whether lipids are released during medium conditioning. The lipid composition of the fat pad conditioned medium was characterized using both thin layer and gas liquid chromatography. These lipid analyses indicated that the fat pad pieces released significant amounts of fatty acids and phospholipids into the medium during the conditioning period. The free fatty acid composition included both saturated and unsaturated molecules, and about 80% of the total fatty acids consisted of palmitate, stearate, oleate, and linoleate. These same fatty acids were a structural component of the majority of phospholipid found in the medium. The addition of palmitate or stearate to defined medium had no effect or was inhibitory for CL-S1 proliferation, depending on the concentration used. Defined medium supplemented with oleate, arachidonate, or linoleate induced CL-S1 proliferation, and the inhibitory effects of palmitate and stearate were overcome by addition of oleate and linoleate. These data indicate that both unsaturated and saturated fatty acids are released from intact adipose cells of the mouse mammary fat pad and that fatty acids can influence the growth of prenoplastic mouse mammary epithelium. Thus, unsaturated fatty acids, perhaps in conjunction with other substances released simultaneously, are candidate molecules for the substances that mediate the effect of adipose tissue on growth of epithelium.

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Author notes

  1. James C. Beck

    Present address: Cell and Molecular Biology Division, Lawrence Berkeley Laboratory, University of California, 04720, Berkeley, CA

  2. Bruce A. Watkins

    Present address: Department of Poultry Science, Virginia Polytechnic Institute and State University, 24061, Blacksburg, VA

Authors and Affiliations

  1. Bio-Regulatory Mechanisms Laboratory, Department of Zoology, Washington State University, 99164, Pullman, Washington

    James C. Beck, Howard L. Hosick & Bruce A. Watkins

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  1. James C. Beck
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  2. Howard L. Hosick
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This work was supported in part by a grant from the American Institute for Cancer Research; grant CA 46885 from the National Institutes of Health, Bethesda, MD; and by State of Washington initiative 171.

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Beck, J.C., Hosick, H.L. & Watkins, B.A. Growth of epithelium from a preneoplastic mammary outgrowth in response to mammary adipose tissue. In Vitro Cell Dev Biol 25, 409–418 (1989). https://doi.org/10.1007/BF02624625

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