BMEBT MS Thesis defense by Shrimathi Venugopalakrishnan
Title:
Evaluation of extracellular vesicles released by 3D spheroids and 2D conventional cell culture of the SKOV3 ovarian cancer cell line
Abstract:
Cancer cells communicate with the surrounding tumor microenvironment and establish a metastatic niche through the secretion of small extracellular vesicles (sEVs), which are referred to as exosomes. These exosomes play a critical role in cancer progression by transferring bioactive cargo, including proteins, lipids, mRNAs, miRNAs, and DNA fragments, to healthy cells, thereby enabling distant organ colonization and metastasis. SKOV3, a widely used human ovarian adenocarcinoma cell line, is traditionally cultured under anchorage-dependent (2D monolayer) conditions; however, suspension culture more faithfully mimics the three-dimensional, non-adherent microenvironment encountered in vivo, giving rise to altered cell signalling, spheroid formation, and distinct exosome release profiles. This study compares the characteristics of exosomes secreted by SKOV3 cells under 2D and 3D conditions, isolated through ultrafiltration and Size Exclusion Chromatography (SEC). Scanning Transmission Electron Microscopy (STEM), Dynamic Light Scattering (DLS), Raman spectroscopy, and Flow Cytometry were used for the characterization. Multivariate analysis across FastGrow and McCoy's media as exosome collection media identified the principal components governing exosome physical properties, while optimized SEC runs demonstrated how Sepharose 2B and 6B columns yield exosomal fractions. Additional investigations included spheroid growth analysis, refractive index analysis, storage effects, Surface-Enhanced Raman Scattering (SERS), and BCA protein estimation in exosomal fractions. This study presents an accessible and reproducible isolation protocol that lowers the technical barrier across laboratory settings, laying the groundwork for non-invasive exosome-based cancer screening.
Textiles 011
Milana Vasudev
508-999-9284
mvasudev@umassd.edu
https://umassd.zoom.us/j/96638190702?pwd=SYKURVzk7JtCHoW0zM4UvgNJCbHVc7.1