Circadian Control of Transcriptional and Metabolic Rhythms in Primary Hepatocytes

Abstract

Isolation of primary hepatocytes and culturing these cells ex vivo provides a powerful platform to model liver physiology in vivo. Primary hepatocytes can be cultured for several days, the circadian clock can be synchronized, and these primary cells can be utilized for functional gene regulation analysis and metabolic studies. In this chapter, we describe detailed methodology for isolation of viable primary hepatocytes, techniques for culturing these cells, methods for synchronization of the circadian clock, transfection and luciferase reporter analysis, as well as glucose production assays as a functional readout of metabolic state.

Keywords: Clock-controlled gene expression; Glucose production assays; Luciferase reporter assays; Primary hepatocyte cell culture; Primary hepatocyte isolation; Synchronization of the circadian clock.