Circadian Control of Transcriptional and Metabolic Rhythms in Primary Hepatocytes.
Circadian Control of Transcriptional and Metabolic Rhythms in Primary Hepatocytes.
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DOI：
10.1007/978-1-0716-2249-0_11
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发表时间：
2022
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原代肝细胞的分离和离体培养这些细胞提供了一个强大的平台，在体内模拟肝脏生理学。原代肝细胞可以培养数天，生物钟可以同步，这些原代细胞可以用于功能基因调控分析和代谢研究。在这一章中，我们描述了详细的方法分离活的原代肝细胞，培养这些细胞的技术，方法同步的昼夜节律钟，转染和荧光素酶报告分析，以及葡萄糖生产测定作为一个功能读出的代谢状态。
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.