Project description:Primary astrocyte cultures were prepared from wildtype and Gde3 knockout mouse brain. Conditioned media was collected, concentrated and analyzed using TMT-MS.
Project description:This study tests whether hepatic steatosis, independent of inflammation, alters the hepatocyte protein secretory profile, and examines whether changes in the secretory products contribute to the development of metabolic dysfunction in other cell types. Mouse hepatocytes were isolated by flow cytometry and cultured in serum-free conditions. Conditioned media was used for LC/MS analysis to compare hepatocytes from chow fed animals to high-fat diet animals with liver steatosis.
Project description:We report single nucleus RNAseq data from the mouse intestinal organoids cultured in quiescent or senescent conditioned media. Analysis revealed changes in cell composition and gene expression caused by SASP factors in senescent conditioned media.
Project description:The interplay between skeletal muscle and bone is primarily mechanical, however, biochemical crosstalk by secreted mediators has recently gained increased attention. The aim of this study was to investigate metabolic effects of conditioned medium from osteoblasts (OB-CM) on myotubes and vice versa. Human skeletal muscle cells incubated with OB-CM showed increased glucose uptake and oxidation, and expression of the glucose transporter (GLUT) 1, while fatty acid uptake and oxidation, and expression of the fatty acid transporter CD36 were decreased. This was supported by proteomic analysis where expression of proteins involved in glucose uptake, glycolytic pathways and TCA cycle were enhanced, and expression of several proteins involved in fatty acid metabolism were reduced. Similar effects on energy metabolism were observed in human bone marrow stromal cells differentiated to osteoblastic cells incubated with conditioned medium from myotubes (SKM-CM), with increased glucose uptake and reduced oleic acid uptake. Proteomic analyses of the two conditioned media revealed many common proteins. Thus, our data may indicate a shift in fuel preference from fatty acid to glucose metabolism in both cell types, induced by conditioned media from the opposite cell type, possibly indicating a more general pattern in communication between these tissues.
Project description:The goal of the present study is to determine the miRNA cargo present in epicardial extracellular vesicles. For that, epicardial cells and human primary epicardial cells were cultured. Conditioned media was isolated from mouse epicardial cells and human primary epicardial cells derived from right atrial appendages in two different states: “cobble” (inactive) cells and “spindle” (active) . RNA from EVs was isolated and sequenced to determine the miRNA content profile.
Project description:TGFβ1 is a profibrotic mediator that contributes to a broad spectrum of pathologies, including pulmonary fibrosis (PF). However, the secretome of TGFβ1-stimulated primary human normal lung (NL) fibroblasts has not been well characterized. Using fluorescent 2-dimensional gel electrophoresis (2D-PAGE) and differential gel electrophoresis (DIGE), we identified 37 differentially secreted proteins in the conditioned media of TGFβ1-activated NL fibroblasts and generated a protein-protein association network of the TGFβ1 secretome using STRING. Functional enrichment revealed that biological processes and pathways characteristics of PF were enriched. Using the DrugBank database, we determined that 32 of the secreted proteins are targets of known experimental, investigational and approved drugs. Additionally, by comparing the TGFβ1 secretome of NL fibroblasts to proteomic biomarkers from biological fluids of systemic sclerosis (SSc) patients, we identified 11 overlapping proteins. Together our data validate the TGFβ1-induced secretome of NL fibroblasts as a valid in vitro model that reflects SSc biomarkers and identifies potential therapeutic targets for SSc-PF.
Project description:RNA was sequenced from differentiated C2C12 mouse myotubes that were treated with S2-013 conditioned media with or without resveratrol in comparison to control media
Project description:Normal human dermal fibroblasts (NHDF) were treated under serum-free conditions with cell culture media conditioned by breast cancer cell lines (SkBr3, MDA-MB-468, T47D) for 72 hours and subjected to gene expression profiling with Illumina platform.
Project description:In this study, 3 biological replicates with 3 technical replicates for the conditioned media (CM) and the whole cell lysates (WCL) of C8-D1A cell line were analyzed using 108 LC-MS/MS runs. Each peptide sample was separated into 6 fractions using stageTip based High-pH fractionation. The peptide samples were analyzed using LC-MS/MS instrumentation consisting of a Nanoflow Easy-nLC 1000 that was connected to a Q Exactive mass spectrometer through a nanoelectrospray ion source. All raw files were processed in MaxQuant, version 1.3.0.5 and the Andromeda search engine against the IPI mouse database (version 3.87, 59 534 entries), containing both forward and reverse proteins sequences, and common contaminants. MS/MS searches for the secretome and the whole-cell proteome were performed with the following parameters: carbamidomethylation as a fixed modification; oxidation of methionine and protein N-terminal acetylation as variable modifications; a 20-ppm first-search tolerance; a 6-ppm main-search tolerance. Minimum peptide length was set to six residues. The false discovery rate for all peptides, PTM sites, and protein identifications was set to 0.01.
Project description:The goal of the experiment was to demonstrate if the overexpression of human-Prune-1 in Triple Negative breast cancer cells induces M2-polarization of macrophages in vitro. For this purpose, murine primary cells from breast tumor developed by Genetically Engineered Mouse Models (GEMMs) of TNBC (i.e., MMTV-Wnt1) and metastatic TNBC overexpressing both human Prune-1 and Wnt1 in mammary gland (i.e., MMTV-Prune-1/Wnt1) were obtained. Conditioned media were collected from these primary cells (1x106 cells) after 24 hours. Murine macrophages (J774A.1 and Raw264.7; 1x106) were starved for six hours and then grown for 48 hours in those conditioned media collected from MMTV-Wnt1 and MMTV-Prune-1/Wnt1 cells. Untreated macrophages were used as negative control for the experiment.