Effect of K7M2 derived extracellular vesicles on gene expression of primary bone marrow derived macrophage
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE205440 | GEO | 2023/12/31
REPOSITORIES: GEO
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Project description:Extracellular vesicles have been shown to be highly enriched in proteins with various functions, including organotropic metastasis. To identify possible protein cargos involved in regulating mestastatic potential of osteosarcoma cell line, we performed quantitative mass spectrometry analysis of EVs from highly metastatic K7M2 cells and poorly metastatic parental K7 cells.
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Project description:RNA-seq was performed on cultured human induced pluripotent cell derived cardiomyocytes (iPSC-CMs). There are four groups, with three samples per group: H1,H2,H3. Negative control. Healthy, normoxic iPSC-CMs D1,D2,D3. Positive control. iPSCs cultured under hypoxic (0-1% O2) for 48h with 2% v/v PBS as vehicle control EV1,EV2,EV3. Treatment 1. iPSCs cultured under hypoxic (0-1% O2) for 48h, with cardiac stromal cell derived extracellular vesicles, provided at a dose of 67ng/µl (2% v/v) EV21, EV23, EV24. Treatment 2 iPSCs cultured under hypoxic (0-1% O2) for 48h, with bone marrow mesenchymal stromal cell (BM-MSC) derived extracellular vesicles, provided at a dose of 67ng/µl (2% v/v) All EVs were isolated from cultured human cells using sequential centrifugation methods. Cells were cultured using commercial EV-depleted FBS to avoid contamination of bovine EVs. EVs were validated for CD81, CD9, ALIX positivity, and visualised by cryoEM. Particle to protein ratios were not different between cardiac and bone marrow EV isolates. Therefore EV doses were standardised so that the same dose by protein (67ng/µl) and EV number (~2,000 EVs per iPSC-CM) were added.
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Project description:Effect of K7M2 derived extracellular vesicles on gene expression of primary bone marrow derived macrophage
| PRJNA845739 | ENA
Project description:Purpose: The goal of this study is to generate NGS-derived transcriptome profiles (RNA-seq) from bone marrow-derived macrophages (BMDMs) exposed to extracellular vesicles (EVs) containing the miR-17 family (miR-17 and miR-93). Methods: BMDMs were isolated from mouse bone and were exposed to lung epithelial Beas2B cell-derived EVs transfected with scrambled control RNAs, miR-17 mimics or miR-93 mimics. Whole transcriptome profiles of the exposed BMDMs (n = 4 biological replicates were pooled) were then generated using the Ion Proton™ System. Results: A wide range of transcript expressions were regulated by the EV-containing miR-17 and/or miR-93 in BMDMs. AC118255.2, Phox2a, 1700066J24Rik, and Gm20412 were effectively downregulated by both MV-miR-17 and MV-miR-93. Conclusions: Our study provides potential target transcripts of EV-containing miR-17/93 in macrophages.
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Project description:Healthy brain function is mediated by several complementary signalling pathways, many of which are driven by extracellular vesicles (EVs). EVs are heterogeneous in both size and cargo and are constitutively released from cells into the extracellular milieu. They are subsequently trafficked to recipient cells, whereupon their entry can modify the cellular phenotype. Here, in order to further analyse the mRNA and protein cargo of neuronal EVs, we isolated EVs by size exclusion chromatography from human induced pluripotent stem cell (iPSC)-derived neurons. Electron microscopy and dynamic light scattering revealed that the isolated EVs had a diameter of 30-100 nm. Transcriptomic and proteomics analyses of the EVs and neurons identified key molecules enriched in the EVs involved in cell surface interaction (integrins and collagens), internalisation pathways (clathrin- and caveolin-dependent), downstream signalling pathways (phospholipases, integrin-linked kinase and MAPKs), and long-term impacts on cellular development and maintenance. Overall, we show that key signalling networks and mechanisms are enriched in EVs isolated from human iPSC-derived neurons.
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Project description:Transcriptional comparison of B16F10 cells with B16F10 cell-derived extracellular vesicles (EVs) to identify transcripts enriched or de-enriched in EVs compared to their donor cells.
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