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.
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Acknowledgments
This work was supported by the National Institutes of Health (NIH) and the National Cancer Institute (NCI) through grants, K22CA212045 and R01CA244519 to SM. In addition, the Masri Laboratory is supported by grants through the Concern Foundation, the V Foundation for Cancer Research, the Cancer Research Coordinating Committee, and the Chao Family Comprehensive Cancer Center at the University of California, Irvine.
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Chun, S.K., Masri, S. (2022). Circadian Control of Transcriptional and Metabolic Rhythms in Primary Hepatocytes. In: Solanas, G., Welz, P.S. (eds) Circadian Regulation. Methods in Molecular Biology, vol 2482. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2249-0_11
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DOI: https://doi.org/10.1007/978-1-0716-2249-0_11
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