Rapid screening of lipid metabolism in monolayer cell cultures.

Summary Monolayer cell cultures grown on coverslips in the presence of radioactive lipid precursors were embedded in silica gel layers for extraction and resolution of the labeled products directly by thin-layer chromatography. The method permits rapid screening of lipid metabolism in tissue cultures with a small number of cells. Supplementary key words phospholipids CHO * ether lipids * neutral lipids . Metabolic studies with cell cultures grown as mono- layers are often difficult because of the large number of flasks and manipulations required. The usual methods have employed detachment of monolayers either by trypsinization or by scraping with a rubber policeman followed by extraction of lipids and sub- sequent analyses. Although useful, these methods re- quire many steps that can be expensive and time- consuming when large numbers of samples are ex- amined. In this report we describe a practical procedure for monitoring lipid metabolism in monolayer cul- tures that is based on techniques that have been described for the extraction and chromatography of lipids from tissue slices (1) and microscopic sec- tions of tissues (2-4) directly on thin-layer chromato- grams. We used cells cultured as monolayers on micro- scope coverslips; the slips were subsequently attached to silica gel layers for extraction and analysis of the labeled products formed. The technique provides not only information on the extent to which labeled pre- cursors are incorporated and on their metabolic fate, but it also permits one to evaluate and select suitable solvents for the quantitative extraction of the labeled products. Materials and Methods