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Exosomal subgroups isolated from MSCs by tangential flow filtration combined with size exclusion chromatography reals distinct molecular properties and functions

ABSTRACT: Exosomal subgroups isolated from MSCs by tangential flow filtration combined with size exclusion chromatography reals distinct molecular properties and functions

PROVIDER: PRJNA1079288 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR28055532_1.fastq.gz	Fastqsanger.gz
	SRR28055532_2.fastq.gz	Fastqsanger.gz
	SRR28055533_1.fastq.gz	Fastqsanger.gz
	SRR28055533_2.fastq.gz	Fastqsanger.gz
	SRR28055534_1.fastq.gz	Fastqsanger.gz

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Exosomal subgroups isolated from MSCs by tangential flow filtration combined with size exclusion chromatography reals distinct molecular properties and functions

Project description:Using a tangential flow filtration and size exclusion chromatography system, the exosomes derived from human umbilical cord mesenchymal stem cells (hUC-MSCs) were purified and separated, resulting in two subgroups with different characterization. These subgroups were named S1-Exos and S2-Exos. The different exosomal subgroups were subjected to proteomic analysis using protein mass spectrometry.

2025-05-06 | PXD050064 | Pride

Exosomal subgroups isolated from MSCs by tangential flow filtration combined with size exclusion chromatography reals distinct molecular properties and functions

Project description:Here, by using TFF-SEC system to separate exosomal subgroups, our study provides an industrial-scale method for the scalable production of MSC exosomes with specific functions. The two exosome subgroups, S1-Exos and S2-Exos, differ in their biogenesis, characterization, and content of cargos, resulting in variations in their biological functions. Although stem cells have been widely used in clinical research and therapy, ineffectiveness or lack of stability still exists (26140604, 26908143). In spite of the well-known advantages of MSCs-Exos in clinical applications, our research introduces a new concept that involves leveraging distinct subpopulations of MSC exosomes for targeted therapeutic interventions in specific diseases.

2025-01-15 | GSE256371 | GEO

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RNASeq of HEK293 suspension cell-derived sEVs treated with a 72 h hyperthermic shift to 40°C versus untreated (37°C) controls

Project description:Experiment aimed at understanding the compositional changes of small extracellular vesicles (sEVs) derived from cells grown under hyperthermic conditions. Human embryonic kidney (HEK293) cells were cultured 48 h under normal conditions (37°C/140 rpm/5% CO2) before being subjected to a 72 h, 40°C temperature shift. sEVs were then isolated from the supernatant via tangential flow filtration (TFF) and size exclusion chromatography (SEC)

2023-08-18 | E-MTAB-13285 | biostudies-arrayexpress

Expression data from CD73 positive adipose derived mesenchymal stem cells (CD73+ AD-MSCs) and CD73 negative adipose derived mesenchymal stem cells (CD73- AD-MSCs) of mouse

Project description:MSCs are a heterogeneous population and the specific population of MSCs may exhibit a selective ability for tissue repair. The aim of our research was to adapt the CD73+ subgroup of adipose derived MSCs (AD-MSCs) for the therapy of myocardial infarction (MI). Our results revealed that CD73+ AD-MSCs played more effective role in the acceleration function of cardiac recovery by promoting angiogenesis in a rat model of MI compared to mixed AD-MSCs and CD73- AD-MSCs. Microarray analysis shows differences between CD73+ and CD73- AD-MSCs when transcription profile of these two subgroups were compared, especially in VEGF pathway.

2021-06-02 | GSE167219 | GEO

Distinct molecular properties and functions of small EV subpopulations isolated from human umbilical cord MSCs using tangential flow filtration combined with size exclusion chromatography.

Project description:As functional derivatives of mesenchymal stem cells (MSCs), small extracellular vesicles (sEVs) have garnered significant attention and application in regenerative medicine. However, the technical limitations for large-scale isolation of sEVs and their heterogeneous nature have added complexity to their applications. It remains unclear if the heterogeneous sEVs represent different aspects of MSCs functions. Here, we provide a method for the large-scale production of sEVs subpopulations derived from human umbilical cord mesenchymal stem cells (HucMSCs), utilizing tangential flow filtration combined with size exclusion chromatography. The resulting subpopulations, S1-sEVs and S2-sEVs, exhibited stable variations in size, membrane-marked proteins, and carrying cargos, thereby displaying distinct functions both in vitro and in animal disease models. S1-sEVs, that highly expressed CD9, HRS and GPC1, demonstrated a greater immunomodulatory impact, while S2-sEVs with enriched expression of CD63 and FLOT1/2 possessed enhanced capacities in promoting cell proliferation and angiogenesis. These discrepancies are attributed to the specific proteins and miRNAs they contain. Further investigation revealed that the two distinct sEVs subpopulations corresponded to different biological processes: the ESCRT pathway (S1-sEVs) and the ESCRT-independent pathway represented by lipid rafts (S2-sEVs). Therefore, we propose the potential for large-scale isolation and purification of sEVs subpopulations from HucMSCs with distinct functions. This approach may provide advantages for targeted therapeutic interventions in various MSC indications.

| S-EPMC11714183 | biostudies-literature

Targeting kidney proximal tubules by protein nanocages via glomerular filtration

Project description:Nature exploits cage-like proteins for a variety of biological purposes from molecular packaging and cargo delivery to catalysis. These cage-like proteins are of immense importance in nanomedicine due to their propensity to self-assemble from simple identical building blocks to highly-ordered architecture and the design flexibility afforded by protein engineering. However, delivery of protein nanocages to the renal tubules remains a major challenge because of the glomerular filtration barrier, which effectively excludes conventional size nanocages. Here we show that DNA-binding Protein from Starved cells (Dps)—the extremely small archaeal antioxidant nanocage—is able to cross the glomerular filtration barrier and is endocytosed by the renal proximal tubules. Using a model of endotoxemia, we present an example of the way in which proximal tubule-selective Dps nanocage can limit the degree of endotoxin-induced kidney injury. This was accomplished by amplifying the endogenous antioxidant property of Dps with addition of a dinuclear manganese cluster. Dps is the first-in-class, protein cage nanoparticle that can be targeted to renal proximal tubules through glomerular filtration. In addition to its therapeutic potential, chemical and genetic engineering of Dps will offer a novel nanoplatform to advance our understanding of the physiology and pathophysiology of glomerular filtration and tubular endocytosis.

2019-05-30 | GSE131922 | GEO

RNA sequence of ISL1-MSCs and Ctrl-MSCs

Project description:RNA sequence of ISL1-MSCs and Ctrl-MSCs

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Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine

Project description:Extracellular vesicles (EVs) have a great potential in clinical applications, however their isolation from different body fluids and their characterisation are not yet optimal or standardised. Here we report the results of examining the performance of ultrafiltration combined with size exclusion chromatography (UF-SEC) to isolate EVs from urine. The results reveal that UF-SEC is a fast method and provides high purity. Furthermore, we introduce asymmetrical-flow field-flow fractionation coupled with a UV detector and multi-angle light-scattering detector (AF4/UV-MALS) as a characterisation method and compare it with the applicabilities and limitations of current methods. We demonstrate that AF4/UV-MALS is a straightforward characterisation method for urinary EVs.

2018-07-04 | PXD009709 | Pride

Exosomes Produced from 3D Cultures of MSCs by Tangential Flow Filtration Show Higher Yield and Improved Activity.

Project description:Exosomes can deliver therapeutic RNAs to neurons. The composition and the safety profile of exosomes depend on the type of the exosome-producing cell. Mesenchymal stem cells are considered to be an attractive cell type for therapeutic exosome production. However, scalable methods to isolate and manufacture exosomes from mesenchymal stem cells are lacking, a limitation to the clinical translation of exosome technology. We evaluate mesenchymal stem cells from different sources and find that umbilical cord-derived mesenchymal stem cells produce the highest exosome yield. To optimize exosome production, we cultivate umbilical cord-derived mesenchymal stem cells in scalable microcarrier-based three-dimensional (3D) cultures. In combination with the conventional differential ultracentrifugation, 3D culture yields 20-fold more exosomes (3D-UC-exosomes) than two-dimensional cultures (2D-UC-exosomes). Tangential flow filtration (TFF) in combination with 3D mesenchymal stem cell cultures further improves the yield of exosomes (3D-TFF-exosomes) 7-fold over 3D-UC-exosomes. 3D-TFF-exosomes are seven times more potent in small interfering RNA (siRNA) transfer to neurons compared with 2D-UC-exosomes. Microcarrier-based 3D culture and TFF allow scalable production of biologically active exosomes from mesenchymal stem cells. These findings lift a major roadblock for the clinical utility of mesenchymal stem cell exosomes.

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