Characterisation of exosomes isolated by ultracentrifugation from human primary bronchial epithelial cells

N. Downey, L. McGarvey, S.L. Martin, A. Crilly, F.T. Lundy

Research output: Contribution to journalMeeting Abstract

Abstract

Introduction:
Exosomes are small extracellular vesicles, approximately 50–150 nm in size, which contain a range of proteins, mRNAs and miRNAs, and have been implicated in cell-cell communication. We have undertaken a study to investigate the functional role of exosomes in cell-cell cross talk between airway epithelial cells and sensory nerves with relevance to cough hypersensitivity in Chronic Obstructive Pulmonary Disease (COPD). Here we report on isolation and characterisation of exosomes from human primary bronchial epithelial cells (PBECs).

Methods:
Exosomes were isolated from cell culture supernatant of PBECs using an ultracentrifugation protocol in line with recommendations from the International Society for Extracellular Vesicles (ISEV) (The´ry et al., 2018). Following isolation, exosomes were characterised using Nanoparticle Tracking Analysis (NanoSight NS300, Malvern Panalytical) to confirm their size, and Transmission Electron Microscopy (TEM) to confirm characteristic morphology.

Results:
Exosomes were successfully isolated from human PBECs and their appropriate size and morphology was confirmed. Nanoparticle Tracking Analysis showed that isolated exosomes had a mean size of approximately 100 nm, a modal size of approximately 80 nm and an estimated concentration of approximately 3.3 9 109 particles/ mL. TEM showed isolated exosomes had a characteristic cup shape and were within the expected size range.

Discussion:
Using an ultracentrifugation protocol we confirm the isolation of exosomes from PBECs, which will be used in subsequent experiments to investigate bronchial epithelial and nerve cell communication and its relevance to heightened cough responses in the COPD airway.
Original languageEnglish
Pages (from-to)36-36
Number of pages1
JournalLung
Volume198
Issue number1
DOIs
Publication statusPublished - 3 Feb 2020
EventAmerican Cough Conference - Reston, United States
Duration: 7 Jun 20198 Jun 2019
Conference number: 7

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