Fluorescence labeling of extracellular vesicles for diverse bio-applications<i>in vitro</i>and<i>in vivo</i>

He, Ying; Xing, Yanlong; Jiang, Tongmeng; Wang, Juan; Sheng-gang, Sang; Rao, Hong; Yu, Fabiao

doi:10.1039/d3cc00998j

Cited by 9 publications

(7 citation statements)

References 119 publications

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“…Fluorescent labeling has the advantages of high stability, sensitivity, and simplicity, and is thus widely used for quantitative analyses of drugs and biomolecules [11]. The introduction of uorescent groups allows differentiation between the labeled polysaccharide and endogenous molecules and thus facilitates in vivo pharmacokinetic research.…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, studies on the pharmacokinetics, tissue distribution, and mechanisms of action of polysaccharides are limited. To address this challenge and promote the in-depth study of P. cyrtonema polysaccharides, the establishment of qualitative and quantitative assays of polysaccharide samples in plasma and tissues is necessary.Fluorescent labeling has the advantages of high stability, sensitivity, and simplicity, and is thus widely used for quantitative analyses of drugs and biomolecules [11]. The introduction of uorescent groups allows differentiation between the labeled polysaccharide and endogenous molecules and thus facilitates in vivo pharmacokinetic research.…”

mentioning

confidence: 99%

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WITHDRAWN: Study on the pharmacokinetics of Polygonatum cyrtonema polysaccharide DPC1 through fluorescence labeling

Yong,

Zhang,

Cao

et al. 2024

Preprint

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Polygonatum cyrtonema is a medicinal plant and its polysaccharides are used for immunomodulation and the treatment of hypoglycemia. Investigation of the tissue distribution and pharmacokinetics of P. cyrtonema polysaccharide can further elucidate its pharmacological mechanism. A fluorescence labeling approach using rhodamine B (RhB) as a fluorescent molecular probe was used for the quantitative assessment of the polysaccharide from dried P. cyrtonema (DPC1) samples, and the pharmacokinetics and tissue distribution of DPC1 were evaluated in mice after intraperitoneal or oral administration. DPC1 was successfully labeled with RhB, showing degrees of fluorescence labeling at 0.453% and 0.568% as determined by the ultraviolet and enzyme marker methods, respectively. DPC1-RhB was rapidly absorbed into the bloodstream after oral and intraperitoneal administration. The relative bioavailability of DPC1-RhB was as high as 48.648%, showing linear pharmacokinetic characteristics. After administration, DPC1-RhB was primarily distributed in the tissues of the heart, spleen, and lung, indicating that the drug has a targeted effect on these tissues. Overall, the findings provide a comprehensive reference for the in vivo distribution of DPC1, together with a foundation for further elucidation of its pharmacological mechanism and the development and application of DPC1 formulations.

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Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

WITHDRAWN: Study on the pharmacokinetics of Polygonatum cyrtonema polysaccharide DPC1 through fluorescence labeling

Yong,

Zhang,

Cao

et al. 2024

Preprint

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“…This, in turn, facilitates the advancement of EV therapeutics and diagnostic applications in clinical settings. 260 Starting from the consideration that each of these EV labelling methods has its strengths and weaknesses (Table 2), therefore, the technique used should reflect the goals of the experiment. Here, the main focus will be on current fluorescent labelling strategies for isolated EVs or direct EV labelling, therefore, this review only briefly provides an overview and examples of the current applications of indirect EV labelling.…”

Section: Methods For Fluorescent (Or Phosphorescent) Labelling Of Ext...mentioning

confidence: 99%

Fluorescent, phosphorescent, magnetic resonance contrast and radioactive tracer labelling of extracellular vesicles

Wardhani,

Levina,

Grau

et al. 2024

Chem. Soc. Rev.

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“…Small EVs labeling methods have been categorized based on the used medium, including fluorescent dyes, fluorescent proteins, and physical labeling methods, which enable labeling in various locations such as the outer membrane, inner membrane, or interior of sEVs. [ 8,9 ] Fluorescent dyes, such as PKH67, anthocyanins and CFSE, are widely applied to label sEVs owing to their advantages of stability, resistance to degradation, widespread availability and simplicity in handling. [ 10 − 12 ] However, the application of lipophilic dyes in studying sEVs is complicated with the disadvantages of unbound dyes, nonspecific binding, and short half‐life.…”

Section: Introductionmentioning

confidence: 99%

The fluorescence mKate2 labeling as a visualizing system for monitoring small extracellular vesicles

Mao,

Lv,

Huo

et al. 2024

Biotechnology Journal

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Small extracellular vesicles (sEVs) are nanosized vesicles enclosed in a lipid membrane released by nearly all cell types. sEVs have been considered as reliable biomarkers for diagnostics and effective carriers. Despite the clear importance of sEV functionality, sEV research faces challenges imposed by the small size and precise imaging of sEVs. Recent advances in live and high‐resolution microscopy, combined with efficient labeling strategies, enable us to investigate the composition and behavior of EVs within living organisms. Here, a modified sEVs was generated with a near infrared fluorescence protein mKate2 using a VSVG viral pseudotyping‐based approach for monitoring sEVs. An observed was made that the mKate2‐tagged protein can be incorporated into the membranes of sEVs without altering their physical properties. In vivo imaging demonstrates that sEVs labeled with mKate2 exhibit excellent brightness and high photostability, allowing the acquisition of long‐term investigation comparable to those achieved with mCherry labeling. Importantly, the mKate2‐tagged sEVs show a low toxicity and exhibit a favorable safety profile. Furthermore, the co‐expression of mKate2 and rabies virus glycoprotein (RVG) peptide on sEVs enables brain‐targeted visualization, suggesting the mKate2 tag does not alter the biodistribution of sEVs. Together, the study presents the mKate2 tag as an efficient tracker for sEVs to monitor tissue‐targeting and biodistribution in vivo.

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Fluorescence labeling of extracellular vesicles for diverse bio-applicationsin vitroandin vivo

Abstract: Extracellular vesicles (EVs) are nanosized vesicles enclosed in lipid membrane that are sustainably released by nearly all cell types. EVs have been deemed as valuable biomarkers for diagnostics and effective...

Cited by 9 publications

References 119 publications

WITHDRAWN: Study on the pharmacokinetics of Polygonatum cyrtonema polysaccharide DPC1 through fluorescence labeling

WITHDRAWN: Study on the pharmacokinetics of Polygonatum cyrtonema polysaccharide DPC1 through fluorescence labeling

Fluorescent, phosphorescent, magnetic resonance contrast and radioactive tracer labelling of extracellular vesicles

The fluorescence mKate2 labeling as a visualizing system for monitoring small extracellular vesicles

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