Faculty mentor/PI email address
pestovdg@rowan.edu
Is your research Teaching and Learning based?
1
Keywords
Extracellular Vesicles, RNA, Centrifugation, Biomarker, A549, Nano Particle
IRB or IACUC Protocol Number
N/A
Date of Presentation
5-6-2026 12:00 AM
Poster Abstract
Extracellular vesicles (EVs) have gained attention as potential diagnostic tools due to their presence in body fluids and their ability to carry nucleic acids, including RNA. EV-derived RNA, particularly ribosomal RNA (rRNA), may reflect cellular stress and disease states, making it a promising biomarker1, 2. This project focuses on optimizing methods to isolate EVs and quantify their RNA content using human cell culture models. Differential ultracentrifugation was used to separate vesicle populations based on size, followed by RNA extraction and quantification using a high-sensitivity assay. Initial experiments using media from three 150 mm culture dishes yielded no detectable RNA, whereas scaling up to six 150 mm dishes resulted in successful isolation and measurable RNA concentrations sufficient for downstream applications. Quantitative analysis demonstrated RNA concentrations ranging from approximately 1.97 to 6.56 ng/µL across different centrifugation fractions, with higher RNA yields observed in lower-speed pellets and decreasing concentrations at higher speeds. This trend is consistent with the removal of larger vesicles and cellular debris at increasing centrifugal forces. Nanoparticle tracking analysis further confirmed enrichment of distinct vesicle populations across fractions. Together, these results highlight the importance of sample input and centrifugation parameters in maximizing EV and RNA recovery. Continued refinement of this methodology may improve consistency and support the use of EV-derived RNA as a biomarker for disease processes such as ischemia/reperfusion injury.
Disciplines
Genetics and Genomics | Medicine and Health Sciences | Neoplasms
Included in
Isolation and Quantification of RNA from Extracellular Vesicles Derived from A549 Human Non-Small Cell Lung Cancer Cells Using Ultracentrifugation
Extracellular vesicles (EVs) have gained attention as potential diagnostic tools due to their presence in body fluids and their ability to carry nucleic acids, including RNA. EV-derived RNA, particularly ribosomal RNA (rRNA), may reflect cellular stress and disease states, making it a promising biomarker1, 2. This project focuses on optimizing methods to isolate EVs and quantify their RNA content using human cell culture models. Differential ultracentrifugation was used to separate vesicle populations based on size, followed by RNA extraction and quantification using a high-sensitivity assay. Initial experiments using media from three 150 mm culture dishes yielded no detectable RNA, whereas scaling up to six 150 mm dishes resulted in successful isolation and measurable RNA concentrations sufficient for downstream applications. Quantitative analysis demonstrated RNA concentrations ranging from approximately 1.97 to 6.56 ng/µL across different centrifugation fractions, with higher RNA yields observed in lower-speed pellets and decreasing concentrations at higher speeds. This trend is consistent with the removal of larger vesicles and cellular debris at increasing centrifugal forces. Nanoparticle tracking analysis further confirmed enrichment of distinct vesicle populations across fractions. Together, these results highlight the importance of sample input and centrifugation parameters in maximizing EV and RNA recovery. Continued refinement of this methodology may improve consistency and support the use of EV-derived RNA as a biomarker for disease processes such as ischemia/reperfusion injury.