A novel sorting method uncovers metabolic heterogeneity between mononucleated and binucleated tetraploid hepatocytes

Our approach leverages Hoechst-area to assess total ploidy and Hoechst-height to differentiate mononucleated and binucleated hepatocytes. Transcriptome analysis comparing these two populations revealed that 1x4n hepatocytes exhibit a broader and more metabolically active gene expression profile. Importantly, these metabolic gene expression programs were independent of liver zonation, a well-known driver of metabolic heterogeneity in hepatocytes.

Our findings uncover a previously underappreciated layer of functional diversity in the liver and provide a new framework for studying the physiological and pathological roles of nuclear configuration in hepatocytes.