Exosomes isolation protocols facts and artifacts for cardiac regeneration

Angelini, Francesco; Ionta,; Rossi, Francesca Maria; Pagano, Francesca; Chimenti, Isotta; Messina, Elisa; Giacomello, Alessandro

doi:10.2741/s464

Cited by 11 publications

(4 citation statements)

References 42 publications

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“…Exosomes, which are small bilayer membrane vesicles, can be extracted from most body fluids, such as urine, saliva and plasma [20]. Lipids, proteins, RNAs (microRNAs, mRNAs and ncRNAs), and DNA are the biological cargos of exosomes secreted by most human cells [21].…”

Section: Discussionmentioning

confidence: 99%

Loss of exosomal LncRNA HCG15 prevents acute myocardial ischemic injury through the NF-κB/p65 and p38 pathways

Chen

et al. 2021

Cell Death Dis

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Exosomes are nanosized bilayer membrane vesicles that may mediate intercellular communication by transporting bioactive molecules, including noncoding RNAs, mRNAs, and proteins. Research has shown that exosomes play an important role in acute myocardial infarction (AMI), but the function and regulation of exosomal long noncoding RNAs (lncRNAs) in AMI are unclear. Thus, RNA sequencing (RNA-Seq) was conducted to investigate the exosomal lncRNA transcriptome from MI patients and identified 65 differentially expressed lncRNAs between the MI and control groups. HCG15, one of the differentially expressed lncRNAs, was verified to have the highest correlation with cTnT by qRT-PCR, and it also contributed to the diagnosis of AMI by receiver operating characteristic (ROC) analysis. Upregulation of HCG15 expression facilitated cardiomyocyte apoptosis and inflammatory cytokine production and inhibited cell proliferation. We also confirmed that HCG15 was mainly wrapped in exosomes from AC16 cardiomyocytes under hypoxia, which contributed to cardiomyocyte apoptosis, the release of inflammatory factors, and inhibition of cell proliferation via the activation of the NF-κB/p65 and p38 pathways, whereas suppressing the expression of HCG15exerted opposite effects, In addition, Overexpression of HCG15 aggravated cardiac IR injury in C57BL/6J mice. This study not only helps elucidate exosomal lncRNA function in AMI pathogenesis but also contributes to the development of novel therapeutic strategies.

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Section: Discussionmentioning

confidence: 99%

Loss of exosomal LncRNA HCG15 prevents acute myocardial ischemic injury through the NF-κB/p65 and p38 pathways

Chen

et al. 2021

Cell Death Dis

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“…To discriminate the biological effects of EVs from the whole PL, we isolated the EVs according to methodological standardized guidelines by high‐speed ultracentrifugation. 64 , 65 Afterwards, we investigated whether EVs may convey haemostatic properties, such as aggregation and pro‐coagulant abilities, which are two key physiological properties exerted by platelets but also reported for EVs. 66 We stimulated platelets of healthy subjects with increasing percentages of EVs (5%, 10% and 20%).…”

Section: Resultsmentioning

confidence: 99%

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

et al. 2022

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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

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“…In order to discriminate the biological effects of EVs from the whole PL, we isolated the EVs according to methodological standardized guidelines by high-speed ultracentrifugation 62,63 . Afterwards, we investigated whether EVs may convey haemostatic properties, such as aggregation and procoagulant abilities, that are two key physiological properties exerted by platelets, but also reported for EVs 64 .…”

Section: Resultsmentioning

confidence: 99%

“Human platelet lysate derived extracellular vesicles enhance angiogenesis through miR-126”

Bordin

Chirivì

Pagano

et al. 2022

Preprint

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Objectivesextracellular vesicles (EVs) represent key biological mediators of several physiological functions within the cell microenvironment. Platelets are the most abundant source of EVs in the blood, contributing to the plastic and versatile behaviour of these cells. Similarly, platelet lysate (PL), the best angiogenic performer of platelet origin and routinely employed 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 contained in PL, by investigating the biological and molecular features related to the angiogenic capacity extrapolated from that possessed by PL.Methodswe tested angiogenic ability and molecular cargo in 3D bioprinted models and by RNA sequencing analysis of PL-derived EVs.Resultsa subset of small vesicles is highly represented in PL. The EVs did not retain aggregation ability, but they preserved a low redox state in endothelial cells and significantly increased the formation of angiogenic tubularly-like structures in 3D endothelial bioprinted constructs. EVs resembled the miRNome profile of PL, mainly enriched of small RNAs and of a high amount of miR-126, the most abundant angiogenic miRNA of platelet origin. The transfer of miR-126 in endothelial cells by EVs after the in vitro inhibition of the endogenous miR-126, restored the angiogenic effects of PL-derived EVs.ConclusionsPL is a biological source of available EVs with angiogenic effects involving a miRNAs-based cargo. These properties can be exploited for targeted molecular/biological manipulation of PL, by potentially developing a product exclusively manufactured of EVs.

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Exosomes isolation protocols facts and artifacts for cardiac regeneration

Cited by 11 publications

References 42 publications

Loss of exosomal LncRNA HCG15 prevents acute myocardial ischemic injury through the NF-κB/p65 and p38 pathways

Loss of exosomal LncRNA HCG15 prevents acute myocardial ischemic injury through the NF-κB/p65 and p38 pathways

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

“Human platelet lysate derived extracellular vesicles enhance angiogenesis through miR-126”

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