University of Louisville

Grade Level at Time of Presentation

Senior

Major

Biology

Minor

Spanish

Institution

University of Louisville

KY House District #

34

KY Senate District #

19

Department

Department of Pharmacology and Toxicology

Abstract

Small extracellular vesicles (sEVs), otherwise known as exosomes, are nanoscale vesicles that cells release to communicate and affect their local and remote microenvironments. Cancer cells specifically use their sEVs to enable tumor survival, including inactivation of anti-tumor macrophage immune responses. For sEV studies in vitro, 2D cells grown in adherent culture are typically used to manufacture sEVs. To more closely simulate in vivo tissues, human hepatocellular carcinoma cells (HCCs) can be grown in a 3D ECM as spheroids. However, the ECM-based method is technically challenging as well as time-consuming since it is incredibly difficult to grow individual spheroids for the purpose of extracting scalable numbers of sEVs for downstream applications. To address this issue, we developed a unique 3D suspension spheroid culture system. 3D suspended spheroid culture is expected to more accurately recapitulate the in vivo 3D scenario minimizing the negative aspects of using 3D matrix-based cell culture for sEV production.

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Development of a 3D HepG2 suspension culture system to enable reducible investigations into 2D vs. 3D HepG2 culture-derived sEV biophysical properties and cancer pathway-related miRNA content

Small extracellular vesicles (sEVs), otherwise known as exosomes, are nanoscale vesicles that cells release to communicate and affect their local and remote microenvironments. Cancer cells specifically use their sEVs to enable tumor survival, including inactivation of anti-tumor macrophage immune responses. For sEV studies in vitro, 2D cells grown in adherent culture are typically used to manufacture sEVs. To more closely simulate in vivo tissues, human hepatocellular carcinoma cells (HCCs) can be grown in a 3D ECM as spheroids. However, the ECM-based method is technically challenging as well as time-consuming since it is incredibly difficult to grow individual spheroids for the purpose of extracting scalable numbers of sEVs for downstream applications. To address this issue, we developed a unique 3D suspension spheroid culture system. 3D suspended spheroid culture is expected to more accurately recapitulate the in vivo 3D scenario minimizing the negative aspects of using 3D matrix-based cell culture for sEV production.

 

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