Details

Autor(en) / Beteiligte

• Kalinec, Gilda M.
• Cohn, Whitaker
• Whitelegge, Julian P.
• Faull, Kym F.
• Kalinec, Federico

Titel

Preliminary Characterization of Extracellular Vesicles From Auditory HEI-OC1 Cells

Ist Teil von

• Annals of otology, rhinology & laryngology, 2019-06, Vol.128 (6_suppl), p.52S-60S

Ort / Verlag

Los Angeles, CA: SAGE Publications

Erscheinungsjahr

2019

Quelle

MEDLINE

Beschreibungen/Notizen

• Objectives: Isolate, purify, and characterize extracellular vesicles (EVs) obtained from auditory HEI-OC1 cells, and evaluate their suitability for intracochlear transport and delivery of pharmacological drugs and/or pro-resolution mediators of acute inflammatory processes. Methods: HEI-OC1 EVs were isolated and purified using the exoEasy Maxi Kit, and their size was evaluated by nanoparticle tracking techniques. Bottom-up proteomics of the EVs, either freshly obtained or stored for up to 4 months at −20°C, was performed by LC-ESI-MS/MS. LC-ESI-MS/MS-MRM was used to measure the loading of dexamethasone inside EVs following co-incubation at room temperature for 1 hour with and without 5 minutes sonication. Results: Routinely, we were able to obtain purified fractions of >2 × 109 EVs/mL, with diameters varying between 50 and 800 nm. Bottom-up proteomics showed that among the most abundant EVs proteins, 19.2% were cytoplasmic, 17.2% were membrane localized, 12.3% were cytosolic, and 14.6% were nucleolar. No significant differences between fresh and stored EVs were detected. Importantly, co-incubation of HEI-OC1 EVs (1 × 108 EVs/mL) with dexamethasone (10 mM) resulted in the incorporation of 10.1 ± 1.9 nM dexamethasone per milliliter of EVs suspension. Conclusions: Altogether, the results suggest that EVs from HEI-OC1 cells could be advantageously used as biological nanocarriers for the delivery of specific molecules and pharmacological drugs into the inner ear.