Response to labeled precursor amino acids, varying cell density, and graded amino acid complement for protein synthesis in mammary cell culture

Effect of labeled precursor amino acids, varying cell densities, graded quantities of amino acid complement, and incubation environment on milk protein synthesis were studied with cultures of mammary cells isolated from Sprague Dawley rats. The essential amino acid complement of Eagle's minimal essential medium was used as base. Protein synthesis, measured by incorporation of labeled lysine, leucine, and phenylalanine, was affected by source of label for the "beta-lactoglobulin fraction" and beta-casein but not alpha-lactalbumin. Cell numbers between 6 X 106 and 6 X 107 per 5 ml of culture medium did not significantly alter rates of synthesis. Increasing amounts of amino acid concentration from one to three-fold increased synthesis of "beta-lactoglobulin fraction" and alpha-lactalbumin regardless of cell population. Response to addition of essential amino acid for "beta-lactoglobulin fraction" synthesis was linear over one to five-fold with 30.9 mug/flask per fold addition (linear regression coefficient; squared correlation = .91). Results were similar for beta-casein synthesis; 25.9 mug/flask and squared correlation = .91. No culturing effects between carbon dioxide and conventional incubators were significant.