Dataset Information

Proteome and secretome of HepaRG cells

ABSTRACT: Primary human hepatocytes, the gold standard for in vitro studies of liver-related functions, suffer from uncertain availability and high variability in cell activity. Over years, a number of alternative hepatic cell lines have lost major liver-like functions, but not HepaRG cells. Therefore, their increasing use worldwide today arouses the need for establishing a reference functional status of differentiated HepaRG cells known as HPR116, which originate from the initial cell bank. Deep proteome and secretome analyses enabled us to show that they nicely express, at levels generally close to those in primary hepatocytes, master regulators of the hepatic phenotype, structural elements that ensure liver-like polarisation and factors supporting their transdifferentiation properties. Their highly differentiated status, mitochondrial functionality, hepatokine secretion ability and response to insulin was verified. Overall, the HepaRG cell system appears as robust surrogate cell system to primary hepatocytes, versatile enough to study not only xenobiotic detoxification but also the control of hepatic energy metabolism, secretory function and disease-related signalling pathways.

INSTRUMENT(S): amaZon ETD, impact HD

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hepatocyte, Liver

SUBMITTER: Fabrice BERTILE

LAB HEAD: Sarah Cianférani

PROVIDER: PXD010304 | Pride | 2019-02-27

REPOSITORIES: Pride

ACCESS DATA

Publications

In-Depth Proteome Analysis Highlights HepaRG Cells as a Versatile Cell System Surrogate for Primary Human Hepatocytes.

Tascher Georg G Burban Audrey A Camus Sandrine S Plumel Marine M Chanon Stéphanie S Le Guevel Remy R Shevchenko Valery V Van Dorsselaer Alain A Lefai Etienne E Guguen-Guillouzo Christiane C Bertile Fabrice F

Cells 20190221 2

Of the hepatic cell lines developed for in vitro studies of hepatic functions as alternatives to primary human hepatocytes, many have lost major liver-like functions, but not HepaRG cells. The increasing use of the latter worldwide raises the need for establishing the reference functional status of early biobanked HepaRG cells. Using deep proteome and secretome analyses, the levels of master regulators of the hepatic phenotype and of the structural elements ensuring biliary polarity were found t ...[more]

PMID: 30795634

Similar Datasets

Project description:Rifampicin and efavirenz are commonly used medicines to treat tuberculosis (rifampicin) and HIV/AIDS (efavirenz). Rifampicin and efavirenz are metabolized in the liver and exposure to these medicines can alter microRNA expression in hepatocytes. Changes in microRNA expression may affect metabolism of these medicines and potentially influence how patient's respond to therapy. It is important to understand changes in microRNA expression in a hepatic cell-based model. Differentiated HepaRG cells were used because mRNA expression and induction of drug metabolizing enzymes are comparable to that of primary human hepatocytes yet differentiated HepaRG cells have an extended lifespan. Differentiated HepaRG cells were obtained from Merck Millipore and cultured as an adherent cell line at a density of 315000 cells/well using collagen I coated 24-well plates. Clinically relevant concentrations of rifampicin and efavirenz were selected as 6.4uM efavirenz (dissolved in 100% dimethyl sulfoxide) and 24.4uM rifampicin (dissolved in 100% dimethyl sulfoxide), while 0.02% dimethyl sulfoxide was used as control to minimize toxicity. HepaRG serum-free induction medium was used to dilute efavirenz and rifampicin. On day 7 since thawing differentiated HepaRG cells, these cells were treated with 6.4uM efavirenz or 24.4uM rifampicin or 0.02% dimethyl sulfoxide for a period of 24 hours. Three biological replicates were available for each treatment condition. Total RNA was isolated from the cells, after treatment, using the Quick-RNATM MiniPrep Kit from Zymo Research Corporation. MicroRNA expression profiling was performed using the TaqMan® OpenArray® Human MicroRNA Panel and QuantStudioTM 12K Flex system. The R/Bioconductor package “Automated Analysis of High-Throughput qPCR Data” were used for data analyses. CT values were used for differential expression analysis and microRNAs with a CT value > 35 was considered undetected. MicroRNAs undetected in any of the replicate samples or microRNAs with AmpScore < 1.24 or CqConf < 0.8 were excluded. Quantile normalization was followed by limma analyses to identify differentially expressed microRNAs for efavirenz versus dimethyl sulfoxide and rifampicin versus dimethyl sulfoxide.

Dataset Information

Proteome and secretome of HepaRG cells

Publications

In-Depth Proteome Analysis Highlights HepaRG Cells as a Versatile Cell System Surrogate for Primary Human Hepatocytes.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure