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Effects of culture condition on small RNA transcriptomes in extracellular vesicles released by human bone marrow stromal cells

ABSTRACT: Effects of culture condition on small RNA transcriptomes in extracellular vesicles released by human bone marrow stromal cells

PROVIDER: PRJNA771408 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR16347582.fastq.gz	Fastqsanger.gz
	SRR16347583.fastq.gz	Fastqsanger.gz
	SRR16347584.fastq.gz	Fastqsanger.gz
	SRR16347585.fastq.gz	Fastqsanger.gz
	SRR16347586.fastq.gz	Fastqsanger.gz

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Effects of culture condition on small RNA transcriptomes in extracellular vesicles released by human bone marrow stromal cells

Project description:Various culture media that can rapidly expand bone marrow stromal cells (BMSCs) are currently available. However, the effects of those culture media on the contents of extracellular vesicles released by bone marrow stromal cells have not been fully understood. Using BMSCs from 6 healthy donors were cultured in two different culture media and characterized the small RNA profiles in extracellular vesicles.

2023-08-08 | GSE185942 | GEO

microRNA profile of control vehicle- and coagulation protease factor VIIa-released endothelial extracellular vesicles

Project description:Coagulation protease factor VIIa (FVIIa) is shown to induce anti-inflammatory and barrier protective effects via endothelial cell protein C receptor (EPCR)-dependent, protease-activated receptor-1 (PAR1)-mediated cell signaling. FVII-EPCR-PAR1 signaling also induces the release of extracellular vesicles from endothelial cells. To obtain clues on whether microRNA (miR) carried out by FVIIa-released EEVs contribute to anti-inflammatory and barrier protective effects, we analyzed miR expression in control- and FVIIa-released EEVs by deep sequencing. These data revealed that several anti-inflammatory miR expression was higher (more than 2-fold) in FVIIa-released EEVs compared to control EEVs, the most predominant being miR10a-5p. The differential expression of miR10a-5p and several other abundant miRs were validated by qRT-PCR. Subsequent in vitro and in vivo experiments showed that miR10a in FVIIa-released EEVs contribute to anti-inflammatory and barrier protective effects.

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Characterization of prostate cancer cell expression profile upon treatment with extracellular vesicles derived from senescent stromal cells in the tumor microenvironment

Project description:Recent advances in next generation sequencing (NGS) has revolutionized system-based analysis of genome-wide expression, cellular pathways and responses. We performed this study to establish the transcriptomic profiling (RNA-seq) of human prostate cancer cells exposed to extracellular vesicles (EVs) released by stromal cells developing a typical senescence-associated secretory phenotype (SASP) in a genotoxic setting.

2020-11-24 | GSE128405 | GEO

Proteomic profile of extracellular vesicles released by breast cancer cells

Project description:The objective of the experiment was to compare the proteomic profile of extracellular vesicles released by breast cancer cells in basal untreated condition (C1, C2, etc) or treated with the adipokine leptin (L1, L2 etc).

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Culture Condition of Bone Marrow Stromal Cells Affects Quantity and Quality of the Extracellular Vesicles.

Project description:Extracellular vesicles (EVs) released by bone marrow stromal cells (BMSCs) have been shown to act as a transporter of bioactive molecules such as RNAs and proteins in the therapeutic actions of BMSCs in various diseases. Although EV therapy holds great promise to be a safer cell-free therapy overcoming issues related to cell therapy, manufacturing processes that offer scalable and reproducible EV production have not been established. Robust and scalable BMSC manufacturing methods have been shown to enhance EV production; however, the effects on EV quality remain less studied. Here, using human BMSCs isolated from nine healthy donors, we examined the effects of high-performance culture media that can rapidly expand BMSCs on EV production and quality in comparison with the conventional culture medium. We found significantly increased EV production from BMSCs cultured in the high-performance media without altering their multipotency and immunophenotypes. RNA sequencing revealed that RNA contents in EVs from high-performance media were significantly reduced with altered profiles of microRNA enriched in those related to cellular growth and proliferation in the pathway analysis. Given that pre-clinical studies at the laboratory scale often use the conventional medium, these findings could account for the discrepancy in outcomes between pre-clinical and clinical studies. Therefore, this study highlights the importance of selecting proper culture conditions for scalable and reproducible EV manufacturing.

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mRNA content of extracellular vesicles, microvesicles and exosomes derived from human H9 embryonic stem cells in culture

Project description:Extracellular vesicles released from cultured H9 cells were fractionated by differential ultracentrifugation and their total RNA were isolated for microarray analysis.

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Project description:Intercellular communication is critical for integrating complex signals in multicellular eukaryotes. Vascular endothelial cells and T lymphocytes closely interact during the recirculation and trans-endothelial migration of T cells. In addition to direct cell-cell contact, we show that T cell derived extracellular vesicles can interact with endothelial cells and modulate their cellular functions. Thrombospondin-1 and its receptor CD47 are expressed on exosomes/ectosomes derived from T cells, and these extracellular vesicles are internalized and modulate signaling in both T cells and endothelial cells. Extracellular vesicles released from cells expressing or lacking CD47 differentially regulate activation of T cells induced by engaging the T cell receptor. Similarly, T cell-derived extracellular vesicles modulate endothelial cell responses to vascular endothelial growth factor and tube formation in a CD47-dependent manner. Uptake of T cell derived extracellular vesicles by recipient endothelial cells globally alters gene expression in a CD47-dependent manner. CD47 also regulates the mRNA content of extracellular vesicles in a manner consistent with some of the resulting alterations in target endothelial cell gene expression. Therefore, the thrombospondin-1 receptor CD47 directly or indirectly regulates intercellular communication mediated by the transfer of extracellular vesicles between vascular cells. HuVEC cells were cocultured with exosomes derived either from Jurkat or JinB8 cells culture media. Each condition was done in triplicate. Also, Exosome RNA from Jurkat or JINB8 cells were compared to each other in triplicate.

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Human iPSC-derived cardiomyocytes under hypoxia/normoxia, treated with bone marrow or cardiac stromal cell extracellular vesicles

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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Human iPSC-derived cardiomyocytes under hypoxia/normoxia, treated with bone marrow or cardiac stromal cell extracellular vesicles

Project description:Effects of cardiac stromal cell extracellular vesicles on human iPSC-derived cardiomyocytes response to hypoxia.

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Expression data of Hematopoietic progenitor and stem cells after 18h of culture with or without extracellular vesicles secreted by AFT stromal cells

Project description:Hematopoietic progenitor and stem cells from bone marrow have been sorted by FACS (LSK, Lineage -, Sca1 + and cKit +) and co-culture during 18h without cytokines with or without extracellular vesicles (EV) secreted by AFT stromal cells. We used microarray to detail the modification of gene expression level of LSK cells after contact with AFT Evs

2017-03-01 | GSE94074 | GEO

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