Thesis
A novel method of exosome loading using direct cell transfection
Master of Science (M.S.), Drexel University
May 2026
DOI:
https://doi.org/10.17918/00011484
Abstract
Exosomes are a class of extracellular vesicles capable of carrying drug cargo including nucleic acids across the blood-brain barrier (BBB). Exosomes have potential for the delivery of nucleic acids but the methods for exosomal loading of nucleic acids require standardization and optimization. We have developed a novel loading method for a specific long noncoding RNA with therapeutic potential, H19. This thesis seeks to examine the novel use of direct cell transfection using Polyethylenimine (PEI) as a method for delivering RNA into exosomes produced by HEK293T (293T) cells, a widely used cell system for the production of therapeutic biologics such as virus particles, proteins, and exosomes. 293T cells were expanded in culture and transfected with varying concentrations of H19 prior to exosomal extraction and RNA isolation to characterize the system's behavior using reverse transcription quantitative polymerase chain reaction (RT-qPCR). The results of this characterization indicate the viability of the method for consistently producing greater than 2-fold enrichment in exosomal H19 and loading H19 across a range of concentrations. The method's sensitivity to minor deviations, however, indicates further refinement is necessary for consistent drug loading. This work constitutes the foundations demonstrating potential viability of a new proposed method for exosomal loading of H19 and other nucleic acids.
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Details
- Title
- A novel method of exosome loading using direct cell transfection
- Creators
- Michael J. Woodburn
- Contributors
- Christian Sell (Advisor)Kara L. Spiller (Advisor)
- Awarding Institution
- Drexel University
- Degree Awarded
- Master of Science (M.S.)
- Publisher
- Drexel University
- Number of pages
- vi, 46 pages
- Resource Type
- Thesis
- Language
- English
- Academic Unit
- School of Biomedical Engineering, Science, and Health Systems (1997-2026); Drexel University
- Other Identifier
- 991022193296904721