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The Direct in Vitro Effect of Insulin-Like Growth Factors (IGFs) on Normal Bovine Mammary Cell Proliferation and Production of IGF Binding Proteins

Monsanto Agricultural Company and Monsanto Corporate Research, Monsanto Company St. Louis, Missouri 63198
Department of Medicine, University of North Carolina Chapel Hill, North Carolina 27599

Address correspondence and requests for reprints to: Michael F. McGrath, Animal Sciences Division, Monsanto Agricultural Company, 700 Chesterfield Village Parkway, Chesterfield, Missouri 63198.

Abstract

Mammary epithelial cells isolated from pregnant, nonlactating heifers were grown in vitro using collagen substrates. Using these systems, the truncated form of insulin growth factor-1 (IGF-1) (des-3-IGF-l), IGF-1, and IGF-2 all stimulated a significant (0.5 to 1 fold) increase in cell proliferation (des-3-IGF-l > IGF-1 > IGF-2). When grown in media containing serum plus IGF-1, normal bovine mammary cells also produced and secreted at least four species of IGF-binding protein (IGFBP) ranging from 21K to 48K (as demonstrated by ligand blot analysis). However, cells grown in serum free media secreted detectable quantities of only 2 major forms of IGFBP of 34K and 48K. Using immunoblot analysis, these proteins were identified as IGFBP-2 and IGFBP-3, respectively. Both proteins were inducible by the addition of IGF to the serum free media (relative potency; IGF-1 > des-3-IGF-1 > IGF-2). Using RIA analysis, bovine mammary cells cultured in the presence of IGF-1 produced 20–25 ng/ml IGFBP-2 compared to control cultures which secrete approximately 1.0 ng/ml. Cells exposed to des-3-IGF-1 produced 40–60% less IGFBP-2 whereas insulin and IGF-2 did not stimulate significant IGFBP-2 production. These data indicate that normal bovine mammary cells secrete IGFBP-2 and IGFBP-3. This secretion is stimulated by IGF-1 and des-3-IGF-1 suggesting a mechanism for regulating local IGF activity.

Received January 30, 1991.

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