Influence of cell culture conditions on diet-induced changes in lymphocyte fatty acid composition.

Abstract

The effect of a range of dietary lipids on the fatty acid composition and membrane fluidity of lymphocytes was investigated. The effects of subsequent culture of these lymphocytes in medium containing autologous serum, foetal calf serum or a serum-free supplement were assessed; this was considered important, since many studies investigating the effects of dietary lipid manipulation on immune function have used protocols involving a variety of cell culture conditions when performing tests of immune function. Weanling Lewis rats were fed for 10 weeks on a low-fat (LF; 20 g/kg) diet or on high fat diets containing 200 g/kg of either hydrogenated coconut oil (HCO), olive oil (OO), safflower oil (SO), evening primrose oil (EPO) or menhaden oil (MO). The fatty acid composition of the phospholipid fractions of lymphocytes from the spleen was altered by dietary lipid manipulation, whereas the fatty acid composition of thymic lymphocytes was not modified significantly. In general, the changes in the fatty acid composition of spleen lymphocytes reflected the fatty acid composition of the diets themselves. Despite the considerable changes in the fatty acid composition of lymphocytes from spleen, dietary lipid manipulation had no effect on the plasma membrane fluidity of these cells. Culturing lymphocytes in autologous serum allowed some, but not all, of the diet-induced changes in fatty acid composition to be maintained. The effects of dietary lipid manipulation were totally reversed when lymphocytes were cultured in FCS. Culturing lymphocytes in serum-free medium not only reversed any effects of dietary manipulation, but also markedly increased the appearance of palmitoleic and oleic acids, at the expense of palmitic and stearic acids, suggesting activation of the delta 9 desaturase when these cells were cultured in the absence of exogenous lipid. This study suggests that cell culture conditions have significant influence on the changes in lymphocyte fatty acid composition brought about by dietary lipid manipulation and may therefore influence the outcome of functional tests.