Cholesterol ester concentrations and the ester hydrolases in cultured glioblastoma cells: Effect of lipoprotein depleted serum

Abstract

We have investigated the effects of substituting lipoprotein depleted serum (LPDS) for normal fetal calf serum (FCS) in culture media on cholesterol ester concentrations and the activity of the ester hydrolases in cultured glioblastoma (C-6 glial) cells. Glial cells grown in media supplemented with 10% FCS contained 16-23% of total cholesterol as esterified sterol. Total sterol content of the cells cultured in media supplemented with LPDS was reduced by 55-75% as compared to cells cultured in FCS media and none of this sterol was in esterified form. Cholesterol ester hydrolase activity was maximal at pH values of 4.5 and 6.5 and required Triton X-100 for optimal activity. Cholesterol ester hydrolase activity at pH 4.5 was significantly higher in cells grown in FCS media than in cells cultured in LPDS media, but the activity at pH 6.5 was not significantly different. The protein: DNA ratio of cells cultured in FCS was higher than in cells cultured in LPDS. These findings indicate that the increase in cholesterol ester concentrations in cells is accompanied by increased activity of lysosomal cholesterol ester hydrolase; and suggest that, in cells cultured in FCS, the availability of free cholesterol for incorporation into cellular membranes is regulated by cholesterol ester hydrolase. The findings also indicate that changes in growth and differentiation of cells cultured in LPDS may be related to reduced availability of exogenous cholesterol.