Dataset Information

Strategies to Enhance Extracellular Vesicle Production.

ABSTRACT: Extracellular vesicles (EVs) are sub-micrometer lipid vesicles secreted from parental cells with their information such as DNA, RNA, and proteins. EVs can deliver their cargo to recipient cells and regulate the signaling pathway of the recipient cells to determine their destiny. Depending on the cargo of EVs, the recipient cells can be changed into abnormal state or be relieved from diseases. Therefore, EVs has been spotlighted as emerging therapeutics in biomedical research. However, slow EV secretion rate is the major limitation for the clinical applications of EVs. EV secretion is highly environmental dependent and can be regulated by various stimulants such as chemicals, oxygen levels, pH, radiation, starvation, and culture methods. To overcome the limitation of low productivity of EVs, EV stimulation methods have been widely studied and applied to massive EV productions. Another strategy is the synthesis of artificial EVs from cells by physical methods such as nitrogen cavitation, extrusion via porous membrane, and sonication. These physical methods disrupt cellular membrane and reassemble the membrane to lipid vesicles containing proteins or drugs. In this review, we will focus on how EV generation can be enhanced and recent advances in large scale EV generation strategies.

SUBMITTER: Hahm J

PROVIDER: S-EPMC8325751 | biostudies-literature | 2021 Aug

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Strategies to Enhance Extracellular Vesicle Production.

Hahm Juhee J Kim Jonghoon J Park Jongmin J

Tissue engineering and regenerative medicine 20210718 4

Extracellular vesicles (EVs) are sub-micrometer lipid vesicles secreted from parental cells with their information such as DNA, RNA, and proteins. EVs can deliver their cargo to recipient cells and regulate the signaling pathway of the recipient cells to determine their destiny. Depending on the cargo of EVs, the recipient cells can be changed into abnormal state or be relieved from diseases. Therefore, EVs has been spotlighted as emerging therapeutics in biomedical research. However, slow EV se ...[more]

PMID: 34275103

Similar Datasets

Project description:Probiotic extracellular vesicles are biochemically active structures responsible for biological effects elicited by probiotic bacteria. Lactobacillus spp., which are abundant in the human body (e.g., gut), are known to have anti-inflammatory and antimicrobial properties, and are commonly used in food products, supplements, and in discovery research. There is increasing evidence that Lactobacillus-derived extracellular vesicles (LREVs) have potent immunomodulatory capacity that is superior to probiotics themselves. However, key mechanistic insights into the process that controls production and thus, the function of LREVs, are lacking. Currently, it is unknown how the probiotic culture microenvironment orchestrates the type, yield and function of LREVs. Here, we investigated how multifactor modulation of the biomanufacturing process controls the yield and biological functionality of the LREVs. To achieve this, we selected Lacticaseibacillus rhamnosus as the candidate probiotic, initially cultivated under traditional culture conditions, i.e., 100% broth concentration and pH 5.5. Subsequently, we systematically modified the culture conditions of the probiotic by adjusting three critical process parameters: (1) culture medium pH (pH 3.5, 5.5 and 7.5), (2) growth time (48 and 72 h), and (3) broth concentration (50% and 10% of original broth concentration). EVs were then isolated separately from each condition. The critical quality attributes (CQA) of LREVs, including physical characteristics (size, distribution, concentration) and biological composition (protein, carbohydrate, lipid), were analysed. Functional impacts of LREVs on human epidermal keratinocytes and Staphylococcus aureus were also assessed as CQA. Our findings show that the production of LREVs is influenced by environmental stresses induced by the culture conditions. Factors like broth concentration, pH levels, and growth time significantly impact stress levels in L. rhamnosus, affecting both the production and composition of LREVs. Additionally, we have observed that LREVs are non-toxicity for keratinocytes, the major cell type of the epidermis, and possess antimicrobial properties against S. aureus, a common human skin pathogen. These properties are prerequisites for the potential application of EVs to treat skin conditions, including infected wounds. However, the functionality of LREVs depends on the culture conditions and stress levels experienced by L. rhamnosus during production. Understanding this relationship between the culture microenvironment, probiotic stress response, and LREV characteristics, can lead to the biomanufacturing of customised probiotic-derived EVs for various medical and industrial applications.

Project description:AbstractExtracellular vesicles (EVs) are nano-sized membrane-bound particles containing biologically active cargo molecules. The production and molecular composition of EVs reflect the physiological state of parent cells, and once released into the circulation, they exert pleiotropic functions via transferring cargo contents. Thus, circulating EVs not only serve as biomarkers, but also mediators in disease processes or injury responses. In the present study, we performed a comprehensive analysis of plasma EVs from burn patients and healthy subjects, characterizing their size distribution, concentration, temporal changes, cell origins, and cargo protein contents. Our results indicated that burn injury induced a significant increase in circulating EVs, the response peaked at the time of admission and declined over the course of recovery. Importantly, EV production correlated with injury severity, as indicated by the total body surface area and depth of burn, requirement for critical care/ICU stay, hospitalization length, wound infection, and concurrence of sepsis. Burn patients with inhalation injury showed a higher level of EVs than those without inhalation injury. We also evaluated patient demographics (age and sex) and pre-existing conditions (hypertension, obesity, and smoking) and found no significant correlation between these conditions and overall EV production. At the molecular level, flow cytometric analysis showed that the burn-induced EVs were largely derived from leukocytes and endothelial cells (ECs), which are known to be activated postburn. Additionally, a high level of zona-occludens-1 (ZO-1), a major constituent of tight junctions, was identified in burn EV cargos, indicative of injury in tissues that form barriers via tight junctions. Moreover, when applied to endothelial cell monolayers, burn EVs caused significant barrier dysfunction, characterized by decreased transcellular barrier resistance and disrupted cell-cell junction continuity. Taken together, these data suggest that burn injury promotes the production of EVs containing unique cargo proteins in a time-dependent manner; the response correlates with injury severity and worsened clinical outcomes. Functionally, burn EVs serve as a potent mediator capable of reducing endothelial barrier resistance and impairing junction integrity, a pathophysiological process underlying burn-associated tissue dysfunction. Thus, further in-depth characterization of circulating EVs will contribute to the development of new prognostic tools or therapeutic targets for advanced burn care.

Dataset Information

Strategies to Enhance Extracellular Vesicle Production.

Publications

Strategies to Enhance Extracellular Vesicle Production.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets