Project description:Early during culture of primary mouse HSCs gene expression changes. These expression alterations can be affected by treating cells with histone deacetylase inhibitor, valproic acid Primary mouse Hepatic stellate cells were cultured for short periods of time (4-16-64h) in presence or absence of valproic acid. Gene expression analysis (mouse Gene 1.0 ST arrays according to manufacturerM-bM-^@M-^Ys manual 701880Rev4 (Affymetrix, Santa Clara, CA)), in vitro stellate cell activation and inhibition of the activation by valproic acid treatment.
Project description:Purpose: The goal of this study was to determine biological consequences during liver regeneration following partial hepatectomy in mice by next-generation sequencing. A particular interest was to compare mice with either a floxed b-PDGFR allele to mice that harbored a deletion of b-PDGFR in hepatic stellate cells (HSCs), by crossing b-PDGFR fl/fl mice with transgenic GFAP-Cre mice. Methods: b-PDGFR fl/fl mice or mice with a HSC-specific deletion of b-PDGFR underwent either sham operation or 70% partial hepatectomy. Following 72 hours, livers were collected and total RNA was extracted using tizol, followed by a purification using Quiagen spin columns including an on-column DNAse digestion step. Conclusion: Our study represents a detailed analysis of hepatic transcriptome, with biologic replicates, generated by RNA-seq technology of livers following sham operation or partial hepatectomy in b-PDGFR fl/fl mice or b-PDGFRfl/fl/GRAP-Cre mice. Whole liver mRNA profiles of sham operated livers or livers collected 72hours after partial hepatectomy of beta-PDGFR fl/fl and beta-PDGFR fl/fl/GFAP-Cre (creating a hepatic stellate cell-specific deletion of b-PDGFR) mice were generated by deep sequencing, in duplicate, using Illumina HiSeq2000.
Project description:To investigate the role of Tet2 deficient immune cells in hepatic stellate cell activation, wild type or Tet2 deficient B cells, T cells, and hepatic macrophages were isolated and co-cultured with purified hepatic stellate cells. Gene expression profiling analysis of bulk hepatic stellate cell RNA was then performed.
Project description:Transcriptional alterations during the first hours of hepatic stellate cell activation were determined using RNA Sequencing. Cultured hepatic stellate cells were collected for RNA Sequencing at several timepoints during the first 24 hours. Illumina NovaSeq SP was used for sequencing.
Project description:Gene expression was determined in primary rat hepatic stellate cells during the in vitro activation process in freshly isolated (4h), quiescent (1d), early activated (3d) and fully activated (7d) hepatic stellate cells. The cells were isolated from the liver using density centrifugation and cultured on plastic in DMEM containing serum for the indicated time. RNA was isolated using the Qiagen Rneasy Mini Kit. The Affymetrix Gene Chip Rat Gene 2.0 ST was used for gene expression analysis performed by the genomic core facility of the EMBL (Heidelberg, Germany). All experiments were performed three times with independent animals.
Project description:Transcriptional profiling of human iPS-HSCs overexpressing LHX2 compared with control iPS-HSCs, which were cocultured with human induced pluripotent stem cell-derived hepatic progenitor cells (iPS-HPCs).