Assay method for measurement of net cholesterol flux

A method for determining net cholesterol flux from cholesterol containing cells following exposure to a cholesterol acceptor, comprising the steps of:

• a) providing detectably labeled cholesterol containing cells;
• b) determining a first specific activity for said cholesterol containing cells;
• c) exposing said cholesterol containing cells to a cholesterol acceptor and at least one cholesterol synthesis inhibitor for a period of time sufficient enough to allow cholesterol flux to occur;
• d) determining cholesterol efflux by measuring the amount of detectably labeled cholesterol in the media;
• e) measuring the cellular cholesterol mass and determining a second specific activity;
• f) determining cholesterol influx from the reduction in said second specific activity compared to said first specific activity;
• and
• g) comparing said cholesterol efflux to said cholesterol influx, wherein a net cholesterol efflux is present when cholesterol efflux is greater than the cholesterol influx and wherein a net cholesterol influx is present when cholesterol influx is greater than the cholesterol efflux.