Characterizing the cellular uptake of neural stem-cell derived exosomes using live-cell imaging techniques

Jones, Samuel, Caws, Thomas, Hayes, Anthony, Marsh Durban, Victoria, Corteling, Randolph and Watson, Peter ORCID: https://orcid.org/0000-0003-0250-7852 2019. Characterizing the cellular uptake of neural stem-cell derived exosomes using live-cell imaging techniques. Presented at: ISEV2019 Annual Meeting, Kyoto, Japan, 24-28 April 2019. Journal of Extracellular Vesicles. , vol.8 (sup1) Wiley, IP12. 10.1080/20013078.2019.1593587

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Abstract

Introduction: Neural stem cell derived exosomes (“ExoPr0”); purified from the conditioned medium of a GMP manufactured, conditionally-immortalized human neural stem cell line (“CTX0E03”), demonstrates a unique biodistribution profile in mice compared to exosomes derived from a control producer cell line. We have previously shown that ExoPr0 is able to cross the blood brain barrier, and to further explicate these findings, we investigated the uptake of ExoPr0 at the cellular level using live-cell imaging techniques. Methods: We employed live-cell confocal microscopy to directly visualize uptake of fluorescently labelled exosomes. A quantitative image analysis protocol was developed and applied to assess the uptake of exosomes in a number of cell types. Results: Time course incubations of cells treated with ExoPr0 produced data that revealed heterogeneity in uptake between cell types. ExoPr0 was compared to exosomes derived from a control producer cell line, highlighting source-specific differences in uptake kinetics. Uptake was observed to occur through more than one pathway resulting in trafficking through endo-lysosomal compartments. The effect of cell cycle on the uptake of ExoPr0 was investigated, but was not observed as having a significant influence. Summary/conclusion: Findings from this study have eluded to the specificity of ExoPr0 towards different cell types and work is ongoing to further elucidate the delivery mechanism of ExoPr0 and understand the subcellular trafficking in recipient cells.

Item Type:	Conference or Workshop Item (Paper)
Date Type:	Published Online
Status:	Published
Schools:	Biosciences Pharmacy
Publisher:	Wiley
Last Modified:	31 May 2024 21:19
URI:	https://orca.cardiff.ac.uk/id/eprint/163176

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