Human platelet lysate‐derived extracellular vesicles enhance angiogenesis through miR‐126

Abstract

Objectives: Extracellular vesicles (EVs) are key biological mediators of several physiological functions within the cell microenvironment. Platelets are the most abundant source of EVs in the blood. Similarly, platelet lysate (PL), the best platelet derivative and angiogenic performer for regenerative purposes, is enriched of EVs, but their role is still too poorly discovered to be suitably exploited. Here, we explored the contribution of the EVs in PL, by investigating the angiogenic features extrapolated from that possessed by PL.Methods: We tested angiogenic ability and molecular cargo in 3D bioprinted models and by RNA sequencing analysis of PL-derived EVs.Results: A subset of small vesicles is highly represented in PL. The EVs do not retain aggregation ability, preserving a low redox state in human umbilical vein endothelial cells (HUVECs) and increasing the angiogenic tubularly-like structures in 3D endothelial bioprinted constructs. EVs resembled the miRNome profile of PL, mainly enriched

Remote Adipose Tissue-Derived Stromal Cells of Patients with Lung Adenocarcinoma Generate a Similar Malignant Microenvironment of the Lung Stromal Counterpart

“…A fold change threshold of >1.2 was considered as upregulated [ 32 ]. The analysis for known protein interaction was performed on commonly upregulated cytokines between A- and LMSC by using the STRING software ( https://string-db.org/ , version 11.5) [ 33 ], building the whole network according to the high confidence setting (0.7) and default options. The pathway and process enrichment analysis were performed using Metascape as already described elsewhere [ 34 ].…”

“…Syn-cel-miR-39 spike in synthetic RNA (Qiagen) was added to monitor extraction efficiency. Afterward, on both tissue sections and sera samples the reverse transcription was performed using the “MiRCURY LNA Reverse Transcription Kit” (QIAGEN) in ThermoMixer 5436 (Eppendorf, Italy) according to the following protocol: 42°C for 60 seconds, 95°C for 5 minutes, 4°C forever [ 33 ].…”

“…Selected miRNA levels such as hsa-miR-101, hsa-miR-126-3p, hsa-miR-486, and hsa-Let-7g were quantified by relative quantification using Qiagen LNA-based SYBR green detection method (miRCURY LNA miRNA PCR assay- Qiagen). Briefly, 3 μ l of cDNA was used on the Applied Biosystems 7900HT machine, adding the relevant ROX concentration to the qPCR mix [ 33 ]. The relative miRNA expression was calculated using hsa-miR-16 as endogenous control with the 2-ΔCt method for cancer tissues and miR-16/miR-39 for sera [ 33 ].…”

“…Briefly, 3 μ l of cDNA was used on the Applied Biosystems 7900HT machine, adding the relevant ROX concentration to the qPCR mix [ 33 ]. The relative miRNA expression was calculated using hsa-miR-16 as endogenous control with the 2-ΔCt method for cancer tissues and miR-16/miR-39 for sera [ 33 ].…”

Next-Generation Sequencing Comparative Analysis of DNA Mutations between Blood-Derived Extracellular Vesicles and Matched Cancer Tissue in Patients with Grade 4 Glioblastoma

“…Extracellular vesicles (EVs) have been found in biofluids and represent an intercellular mechanism of biomolecule transport [ 8 , 9 ], including by lipids [ 10 ], proteins [ 11 ], and nucleic acids [ 12 ]. Glioblastoma cells release EVs in the local microenvironment and can be detected in the bloodstream (they are able to cross the blood–brain barrier, which is often compromised in these patients), mediating angiogenesis, proliferation, immunomodulation, carcinogenesis, and invasiveness [ 13 , 14 , 15 ].…”