Project description:Primary hepatocytes are a commonly used in vitro model for studying liver metabolism, but prolonged culturing results in dedifferentiation. Here we characterized the transcriptome and proteome of whole liver and primary hepatocytes as either freshly isolated cells or after 24 hours of 2D-culturing. We found that after 24 hours of 2D-culturing, hepatocytes showed differential expression of over 10,000 genes and 3,000 proteins compared to freshly isolated cells. This was accompanied by a reduced transcriptional heterogeneity and a loss of zonal markers. Cultures contained ~10% non-hepatocytes, including stellate- and dedifferentiated endothelial cells. Culturing also increased abundance of proteins associated with oxidative stress and inflammation, and changed mitochondrial, extracellular, and ribosomal protein abundance. The data generated are available in the shiny-app “Hepamorphosis”. Our finding show that primary mouse hepatocytes undergo significant changes in culture, limiting theirutility for studying physiological and molecular mechanisms related to the liver.
Project description:RNA-seq analysis of primary hepatocytes isolated from C57BL/6N mice, stimulated with IL-1β recombinant protein to examine the transcriptional response to inflammatory signaling.
Project description:This study provides an evaluation of changes in gene expression associated with acetominophen treatment of rat hepatocytes in vitro. Primary rat hepatocytes were treated for 24 and 48 hours with two doses (500 uM and 5 mM) of acetominophen and 1% DMSO vehicle control. Five replicates of each treatment were performed. Cells were then extracted and RNA processed for microarray analysis.
Project description:This study provides an evaluation of changes in gene expression associated with clofibrate treatment of rat hepatocytes in vitro. Primary rat hepatocytes were treated for 24 and 48 hours with two doses (60 uM and 1 mM) of clobibrate and 1% DMSO vehicle control. Five replicates of each treatment were performed. Cells were then extracted and RNA processed for microarray analysis.
Project description:This study provides an evaluation of changes in gene expression associated with phenobarbital treatment of rat hepatocytes in vitro. Primary rat hepatocytes were treated for 24 and 48 hours with two doses (300 uM and 3 mM) of phenobarbital and water vehicle control. Five replicates of each treatment were performed. Cells were then extracted and RNA processed for microarray analysis.
Project description:This study provides an evaluation of changes in gene expression associated with methapyrilene treatment of rat hepatocytes in vitro. Primary rat hepatocytes were treated for 24 and 48 hours with two doses (3 uM and 100 uM) of methaphyriline and 1% DMSO vehicle control. Five replicates of each treatment were performed. Cells were then extracted and RNA processed for microarray analysis.