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Hepatocyte size fractionation allows dissection of human liver zonation
Author(s) -
Ölander Magnus,
Wegler Christine,
Flörkemeier Inken,
Treyer Andrea,
Handin Niklas,
Pedersen Jenny M.,
Vildhede Anna,
Mateus André,
LeCluyse Edward L.,
Urdzik Jozef,
Artursson Per
Publication year - 2021
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.30273
Subject(s) - hepatocyte , fractionation , biology , elutriation , in vitro , in vivo , function (biology) , cell size , cell fractionation , human liver , gene expression , microbiology and biotechnology , gene , biochemistry , chemistry , enzyme , genetics , chromatography , organic chemistry
Human hepatocytes show marked differences in cell size, gene expression, and function throughout the liver lobules, an arrangement termed liver zonation. However, it is not clear if these zonal size differences, and the associated phenotypic differences, are retained in isolated human hepatocytes, the “gold standard” for in vitro studies of human liver function. Here, we therefore explored size differences among isolated human hepatocytes and investigated whether separation by size can be used to study liver zonation in vitro. We used counterflow centrifugal elutriation to separate cells into different size fractions and analyzed them with label‐free quantitative proteomics, which revealed an enrichment of 151 and 758 proteins (out of 5163) in small and large hepatocytes, respectively. Further analysis showed that protein abundances in different hepatocyte size fractions recapitulated the in vivo expression patterns of previously described zonal markers and biological processes. We also found that the expression of zone‐specific cytochrome P450 enzymes correlated with their metabolic activity in the different fractions. In summary, our results show that differences in hepatocyte size matches zonal expression patterns, and that our size fractionation approach can be used to study zone‐specific liver functions in vitro.