RNA-seq of human BM-derived mesenchymal stromal cells (MSCs) cocultured with mouse BM cells upon GM-CSF stimulation against MSCs without BM cell coculture
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ABSTRACT: RNA-seq of human BM-derived mesenchymal stromal cells (MSCs) cocultured with mouse BM cells upon GM-CSF stimulation against MSCs without BM cell coculture
Project description:CD11bloLy6CloLy6Glo cells were sorted from the steady-state bone marrow (BM) of B6 mice, i.e. cells cultured for 5 d without human BM-derived mesenchymal stromal cells (MSCs) in the absence of GM-CSF. CD11bhiLy6ChiLy6Glo cells were isolated from BM cells cultured for 5 d under GM-CSF incubation (40 ng/ml) but without MSC coculture. CD11bmidLy6CmidLy6Glo cells were sorted from GM-CSF-stimulated, MSC-cocultured BM cells.
Project description:In bone marrow (BM), there are two different types of stem/progenitor cells. With respect to hematopoiesis, hematopoietic stem/progenitor cells (HPCs) produce mature blood cells and mesenchymal stromal/stem cells (MSCs) support this. The influence of exposure to low-dose radiation on human HPCs has been investigated, and generation of both immature and mature hematopoietic cells from human HPCs is compromised. On the other hand, the influence of exposure to low-dose radiation on MSCs is not known. This gene expression profiling was created for investigation how low-dose irradiation affects BM-MSCs genomically.
Project description:Human MSCs were cocultured for 18 hours with macrophages which had been differentiated from human peripheral blood-derived monocytes by PMA and sequentially stimulated by 2 μg/mL LPS (InvivoGen) for 3 hours and 5 mM ATP (InvivoGen) for 45 minutes. Normal MSCs without macrophage coculture and MSCs cocultured with activated macrophages were assayed by miRNA arrays.
Project description:Bone marrow-derived multipotent stromal cells (BM-MSCs) exhibit therapuetically valuable properties, including the capacity to differentiate into skeletal tissues and modulate immune system activity. These properties depend on proper regulation of dynamic gene expression in response to environmental and developmental stimuli. This study used chromatin immunoprecipitation (ChIP) coupled with human promoter tiling microarray analysis (ChIP-on-chip) to profile histones H3K4me3 and H3K27me3 at promoters genome-wide. The goal of the study was to identify gene promoters marked by H3K27me3 and H3K4me3 in BM-MSCs.
Project description:BRD4, a member of the BET family of histone readers, binds to acetylated lysine of histone H3 and promotes assembly of super-enhancer complexes that drive expression of key oncogenes in acute myeloid leukemia (AML) and other cancers. ARV-825 is a proteolysis-targeting chimera (PROTAC) that targets BRD4 for CRBN-mediated ubiquitination and degradation. BM-MSCs are an important element of the bone marrow microenvironment of AML. To understand how targeting BRD4 in BM-MSCs may contribute to the overall effect on AML of targeting BRD4, we treated BM-MSCs from two normal donors with ARV-825 in vitro. Treatment of BM-MSC monocultures with ARV-825 for 24 hr caused extensive changes in gene expression, highly uniform between triplicates. Although the cultures from the two normal donors showed different profiles, their changes with ARV-825 were highly similar. These changes implicated effects on oxidative stress, osteogenic differentiation, retinoid metabolism, F-actin polymerization, CXCL12, and proliferation.
Project description:Propionate accumulation is an important bottleneck for anaerobic degradation of organic matter. We hypothesized that propionate conversion by a novel coculture of Syntrophobacter fumaroxidans and Geobacter sulfurreducens can be an alternative strategy for propionate oxidation coupled to Fe(III) reduction. In this study, we successfully cocultured S. fumaroxidans and G. sulfurreducens on propionate and Fe(III). Proteomic analyses of this coculture provided insights into the underlying mechanisms of propionate metabolism pathway and interspecies electron transfer mechanism. Our study can be further useful in understanding syntrophic propionate degradation in bioelectrochemical and anaerobic digestion systems.
Project description:Analysis of genes induced in DC precursors and in BM cells and monocytes treated with GM-CSF For progenitor arrays, bone marrow progenitors (CMP, GMP, CDP, and pre-cDC) were harvested from WT C57Bl/6 mice. For culture arrays, BM was cultured in the presence of GM-CSF or M-CSF and adherent and non-adherent cells sorted. For monocyte cultures, sorted BM monocytes were treated with GM-CSF for 0, 24 or 48 hours.