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Real-time quantitative PCR analysis of human mesenchymal stromal (MSC) cells extracellular vesicles (EV)

ABSTRACT: Real-time quantitative PCR analysis of human mesenchymal stromal (MSC) cells extracellular vesicles (EV)

PROVIDER: PRJNA659509 | ENA |

REPOSITORIES: ENA

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Real-time quantitative PCR analysis of human mesenchymal stromal (MSC) cells extracellular vesicles (EV)

Project description:Bone Marrow MSC were isolated from 5 donnors, EV were isolated by unltracentrifugation from the conditionned media after 48h without serum. 3 conditions are described: control, interferon priming (25 ng/ml) or hypoxia priming (O2 = 3%)

2020-08-27 | GSE156919 | GEO

MicroRNA expression profiling of extracellular vesicles derived from human BM-MSC

Project description:Human MSC are tissue stem cells that show multiple biological effects. At the present, how BM-MSC exert their effects are not fully understood. In this study, a novel cell-cell communication mediator, extraxellular vesicles (EV), were examined in the involvement of BM-MSC-mediated biological effects.

2018-05-21 | GSE99082 | GEO

The incorporation of extracellular vesicles from MSC into HSC increase their clonogenic capacity and engraftment ability

Project description:Treatment of graft failure after allogeneic stem cell transplantation with mesenchymal stromal cells (MSC) is being evaluated clinically. MSC can exert their functions by the release of extracellular vesicles (EV). Our aim is to analyze changes induced in hematopoietic stem cells (HSC) after the incorporation of MSC-EV. MSC were isolated from healthy donors. MSC-EV were isolated by ultracentrifugation and characterized by flow cytometry (FC), Western-blot, electron microscopy and Nanosight. EV incorporation into HSC was analyzed by FC and confocal microscopy. To study HSC gene expression, RT-PCR and arrays were performed. Apoptosis and cell cycle were evaluated by FC and clonal growth by clonogenic assays in methylcellulose. The engraftment capacity of HSC was analyzed four weeks after transplantation in NOD-SCID mice. Our results showed that MSC-EV presented the characteristic morphology, size and inmunophenotype and were able to be incorporated into HSC. MSC-EV incorporation induced an a down-regulation of pro-apoptotic genes, an overexpression of genes involved in colony formation and cell proliferation together with an activation of the JAK-STAT pathway in HSC. A significant decrease in apoptosis (p=0.001), an increase of the percentage of cells in phase S and an increased CD44 expression (p=0.013) were confirmed by FC in HSC with MSC-EV. In addition, these HSC displayed a higher CFU-GM clonogenic potential (p=0.010). Finally, in the xenotransplantation model, the engraftment ability of human HSC with MSC-EV was significantly higher (p= 0.027). In summary, the incorporation of MSC-EV induces genomic and functional changes in HSC, increasing their clonogenic capacity and their engraftment ability.

2020-10-31 | GSE120803 | GEO

Effect of MSC and MSC-derived extracellular vesicles on human blood coagulation

Project description:Mesenchymal stem cells (MSC) have emerged as potent therapeutic tool for a number of pathologies, including immune ones. However, unwelcome effects of MSC on the blood coagulation were revealed in some cases, which require more in-depth analysis. In this study, we explored the trombotic properties of human MSC from umbilical cord. We revealed strong procoagulant effects of umbilical cord MSC toward human and rat whole blood and platelets-free plasma using rotational thromboelastometry and thrombodynamics tests. The similar potentiation of clotting was demonstrated for MSC-derived extracellular vesicles (EV). In order to suggest approaches to avoid unwanted effects we studied the impact of heparin supplement on MSC/EV procoagulation properties. We found that therapeutic doses of unfractionated heparin injected in the patient's blood (administered in vivo) did not abrogate the procoagulant properties of MSC. Mass-spectrometry analysis of proteins of MSC and EV involved in coagulation-associated pathways was used to evaluate mechanisms of protrombotic effects.

2019-03-26 | PXD012768 | Pride

Real-time quantitative PCR analysis of EV-associated miRNAs from Human Tconv cell

Project description:Real-time quantitative PCR analysis of EV-associated miRNAs from Human Tconv cell

| PRJNA761723 | ENA

Real-time quantitative PCR analysis of rat's brain miRNAs

Project description:We investigated the therapeutic effect of the extracellular vesicles secreted by glial progenitor cells (GPC-EV) derived from human induced pluripotent stem cell in a traumatic brain injury model. The injury was modeled using the dosed concussion to the open brain method. The male Wistar rats were performed traumatic brain injury (TBI), and after 24 hours animals were randomly divided into two groups: group 1 — rats with intranasal administration of PBS (control), and group 2 — rats with similar administration of extracellular visicles (EV) derived from glial progenitor cells. miRNA PCR analysis of brain tissues ( cortex, striatum, hippocampus) were performed on postoperative day 7 to investigate miRNA associated with apoptosis, and inflammation.

2023-09-15 | GSE242828 | GEO

Real-time quantitative PCR analysis of rat's brain mRNAs

Project description:We investigated the therapeutic effect of the extracellular vesicles secreted by glial progenitor cells (GPC-EV) derived from human induced pluripotent stem cell in a traumatic brain injury model. The injury was modeled using the dosed concussion to the open brain method. The male Wistar rats were performed traumatic brain injury (TBI), and after 24 hours animals were randomly divided into two groups: group 1 — rats with intranasal administration of PBS (control), and group 2 — rats with similar administration of extracellular visicles (EV) derived from glial progenitor cells. PCR analysis of brain tissues ( cortex, striatum, hippocampus) were performed on postoperative day 7 and 14 to investigate genes associated with apoptosis, and inflammation.

2023-09-13 | GSE242996 | GEO

EV-RNA sequencing with RNA TOP-PCR for healthy males

Project description:RNA sequencing via RNA TOP-PCR of EV from healthy males

2021-06-01 | GSE152693 | GEO

Extracellular vesicles (EV) secreted by embryonic stem cell-derived cardiovascular progenitor cells (hES-CPg) recapitulate the therapeutic effects of these cells in a model of chronic heart failure (CHF).

Project description:Background: We have found that extracellular vesicles (EV) secreted by embryonic stem cell-derived cardiovascular progenitor cells (hES-CPg) recapitulate the therapeutic effects of these cells in a model of chronic heart failure (CHF). Objectives: Our goal was to test other cellular sources of EV and to explore their mechanism of action.

2017-10-06 | E-MTAB-5164 | biostudies-arrayexpress

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.

2024-10-02 | E-MTAB-13966 | biostudies-arrayexpress

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