Cholesterol is a biological molecule of great significance. Along with being a fundamental component of the cell membrane, it is also an important precursor to bile acids, steroid hormones, and vitamin D. Considerable progress has been made in understanding the importance of cholesterol to cells, its mechanism of transport in the body, its complex regulatory pathways, and clinical significance. While novel roles of cholesterol in human physiology and pathology are continually being uncovered, obtaining a better mechanistic understanding of these functions will provide new insight into diseases associated with cholesterol and help to identify therapeutic interventions.