Reply to: Letter to Editor by Saravanan Sampoornam et al.

Journal of Cellular and Molecular Medicine, Journal Year: 2025, Volume and Issue: 29(5)

Published: March 1, 2025

Language: Английский

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Proteomic profiling of tissue extracellular vesicles (EVs) identifies tissue-specific EV markers and predicts the accessibility of tissue EVs to the circulation

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: March 11, 2025

Summary Extracellular vesicles (EVs) mediate cell-to-cell communication in endocrine, paracrine, and autocrine fashions, but their roles transport the body remain incompletely understood. It is expected that EVs circulate through bloodstream, where derived from various tissues are present. However, how much each tissue contributes to circulating largely unknown. Therefore, identifying tissue-specific EV markers of importance for elucidating vivo dynamics EVs. A key issue hampers studying lack methodologies collecting them tissues. Here, we developed methods isolate four different tissues, skeletal muscle, heart, liver, adipose tissue, performed proteomic analysis on Unbiased quantitative revealed protein signatures identified marker proteins specific tissue. Furthermore, comprehensive comparisons proteome plasma EVs, predicted accessibility circulation. Our data suggests that, among tissue-derived more readily enter circulation, while muscle-derived exhibit poor blood. Collectively, our circulation highly depends original

Language: Английский

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Systems Approaches to Cell Culture-Derived Extracellular Vesicles for Acute Kidney Injury Therapy: Prospects and Challenges

Function, Journal Year: 2024, Volume and Issue: 5(3)

Published: Jan. 1, 2024

Abstract Acute kidney injury (AKI) is a heterogeneous syndrome, comprising diverse etiologies of insults that result in high mortality and morbidity if not well managed. Although great efforts have been made to investigate underlying pathogenic mechanisms AKI, there are limited therapeutic strategies available. Extracellular vesicles (EV) membrane-bound secreted by various cell types, which can serve as cell-free therapy through transfer bioactive molecules. In this review, we first overview the AKI syndrome EV biology, with particular focus on technical aspects application culture-derived EVs. Second, illustrate how multi-omic approaches miRNA, protein, genomic cargo analysis yield new insights into their action address unresolved questions field. We then summarize major experimental evidence regarding potential EVs subdivide stem non-stem cell-derived Finally, highlight challenges opportunities related clinical translation animal studies human patients.

Language: Английский

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The effect of culture media and seeding densities on the human malignant testicular germ cell line (NT2/D1) growth pattern

Journal of Biological Research - Bollettino della Società Italiana di Biologia Sperimentale, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 5, 2025

Testicular cancer is a common malignancy in young males, comprising 5% of urological cancers and 1% all male cancers. The NT2/D1 cell line, pluripotent embryonal carcinoma, valuable developmental biology research due to its differentiation potential into neurons glial cells. Although Fetal Bovine Serum (FBS) often used promote growth, limitations necessitate optimized culture conditions. Monitoring growth health through methods such as counting, viability assays, curves, microscopy crucial for reliable research, particularly evaluating specific characteristics before vitro studies. study compared the patterns lines under different conditions seeding densities using trypan blue counting MTT methods. cultured heat-inactivated FBS (H.iFBS) non-heat-inactivated (Non-H.iFBS) exhibit distinct patterns, doubling times, morphologies. Cell density condition medium are two key parameters influencing metabolic activities enhancing variability. There no preference between culturing cells yet it should be considered experiment design. Thus systematic reporting essential facilitate better comparisons vivo experimental data.

Language: Английский

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Engineered dendritic cells-derived extracellular vesicles for cancer immunotherapy

Journal of Controlled Release, Journal Year: 2025, Volume and Issue: unknown, P. 113620 - 113620

Published: March 1, 2025

Language: Английский

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Extracellular vesicles: innovative cell-free solutions for wound repair

Frontiers in Bioengineering and Biotechnology, Journal Year: 2025, Volume and Issue: 13

Published: April 3, 2025

Chronic non-healing wounds are often associated with conditions such as diabetes and peripheral vascular disease, pose significant medical socioeconomic challenges. Cell-based therapies have shown promise in promoting wound healing but major drawbacks immunogenicity tumor formation. As a result, recent research has shifted to the potential of extracellular vesicles (EVs) derived from these cells. EVs nanosized lipid bilayer vesicles, naturally produced by all cell types, which facilitate intercellular communication carry bioactive molecules, offering advantages low immunogenicity, negligible toxicity be re-engineered. Recent evidence recognizes that during released wide range cells including immune cells, skin epithelial platelets they actively participate repair. This review comprehensively summarizes latest on function endogenous types different phases healing, thereby presenting interesting therapeutic targets. Additionally, it gives critical overview current status mesenchymal stem cell-derived treatment highlighting their tremendous non-cellular of-the-shelf alternative care.

Language: Английский

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Rapid isolation of extracellular vesicles from stem cell conditioned medium using osmosis-driven filtration

Science and Technology of Advanced Materials, Journal Year: 2025, Volume and Issue: unknown

Published: April 3, 2025

Extracellular vesicles (EVs) hold significant promise as biomarkers and therapeutics, yet their isolation remains challenging due to low abundance complex sample matrices. Here, we introduce EV-Osmoprocessor (EVOs), a novel device that leverages osmosis-driven filtration for rapid efficient EV isolation. EVOs employs high osmolarity polymer solution concentrate EVs while simultaneously removing smaller contaminants. Compared traditional methods such ultracentrifugation precipitation, offers speed convenience, achieving 50-fold volume reduction in under 2 h. Our results show retained removed >99% albumin from the cell conditioned culture medium (CCM). The isolated exhibited particle size distribution centered around 140 nm, which was very similar via precipitation or ultracentrifugation. standalone process achieved particle:protein ratio (EV purity) of ~107 particles/µg protein. Comprehensive characterization, including cryo-electron microscopy, validation protein markers known miRNA cargo confirmed successful EVs. Functional assays, based on protection cardiomyocytes hypoxia/reoxygenation injury, demonstrated bioactivity EVOs-isolated Furthermore, can be used 30 ml CCM into 0.5 solution, then further processed with size-exclusion chromatography (SEC), improving purity ~109 This work establishes promising tool research clinical applications, offering streamlined approach enhanced analytical performance.

Language: Английский

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Do Media Extracellular Vesicles and Extracellular Vesicles Bound to the Extracellular Matrix Represent Distinct Types of Vesicles?

Biomolecules, Journal Year: 2023, Volume and Issue: 14(1), P. 42 - 42

Published: Dec. 28, 2023

Mineralization-competent cells, including hypertrophic chondrocytes, mature osteoblasts, and osteogenic-differentiated smooth muscle cells secrete media extracellular vesicles (media vesicles) bound to the matrix (matrix vesicles). Media are purified directly from medium. On other hand, after discarding medium subjecting embedded in or bone cartilage tissues an enzymatic treatment. Several pieces of experimental evidence indicated that isolated same types mineralizing have distinct lipid protein composition as well functions. These findings support view released by different functions mineralized due their location, which is anchored versus free-floating.

Language: Английский

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Proteomic Profiling of Mesenchymal Stem Cell-Derived Extracellular Vesicles: Impact of Isolation Methods on Protein Cargo

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 7, 2024

Abstract Extracellular vesicles (EVs) are nanosized with a lipid bilayer that secreted by cells and play critical role in cell-to-cell communication. Despite the promising reports regarding their diagnostic therapeutic potential, utilization of EVs clinical setting is limited due to insufficient information about cargo lack standardization isolation analysis methods. Considering protein cargos as key contributors potency, we conducted tandem mass tag (TMT) quantitative proteomics three subpopulations mesenchymal stem cell (MSC)-derived obtained through different techniques: ultracentrifugation (UC), high-speed centrifugation (HS), on sucrose cushion (SU). Subsequently, checked EV marker expression, size distribution, morphological characterization, followed bioinformatic analysis. The proteome results revealed these exhibit distinct molecular functional characteristics. choice method impacts isolated isolating EVs. Specifically, (HS) exhibited higher abundance ribosomal mitochondrial proteins. Functional apoptosis assays comparing mitochondria methods HS-EVs, but not other EVs, induced early cancer cells. On hand, using (SU) (UC) demonstrated proteins primarily involved immune response, cellLJcell interactions, extracellular matrix interactions. Our analyses unveil notable disparities associated biological functions among subpopulations, underscoring importance meticulously selecting resultant based intended application.

Language: Английский

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Extracellular vesicles derived from stored red blood cell suspensions enhance invasion and migration of lung cancer cells by miR1246 and miR150‐3p

Vox Sanguinis, Journal Year: 2024, Volume and Issue: 119(8), P. 809 - 820

Published: June 5, 2024

Aged red blood cell (RBC) transfusions in lung cancer patients are often related to recurrence and shorter lifespans. Extracellular vesicles (EVs) accumulated stored RBC suspensions may be one of the important influential factors. This study aims investigate how EVs derived from affect progress through most enriched microRNAs (miRNAs) previously reported our research.

Language: Английский

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