Researcher(s)
- Kevin Schmidt, Biological Sciences, University of Delaware
Faculty Mentor(s)
- Arit Ghosh, UD Flow Cytometry Core, University of Delaware
Abstract
As science advances, new technologies arise to analyze cells at a smaller and smaller level. These advancements allow us to study cell processes that were previously impossible, such as cell to cell communication. The communication between cells in a multicellular organism is crucial for survival in their ever changing world. Exosomes are small vesicles released by cells for the purpose of cell to cell communication. The cargo carried by exosomes often alter the fates of the cells receiving. Exosomes are believed to have many potential therapeutic applications and have made significant impact in various fields of biology such as cancer treatment to synthetic biology. This project focuses on studying specific exosomes that are made by blood platelets which are positive for the surface protein – CD63. In order to study CD63 exosomes in an in vitro setting, DNA plasmids were introduced into human embryonic kidney cells (HEK293T) to facilitate the expression of a fluorescent (GFP) version CD63 in these kidney cells, allowing for the analysis and quantification of CD63 expression using flow cytometry. The use of flow cytometry allows us to detect the proteins on a cell-by-cell basis, giving us an accurate representation of the population of cells. However, as CD63 exosomes are extruded into the cellular environment we need to detect these vesicles via a small particle detector due to their size range of 50-150 nm. This has necessitated extensive cleaning, maintenance and quality control operating protocols for a CytoFLEX LX (capable of small particle detection) at the UD Flow Cytometry Core. We plan to connect the results of this project into a future study using platelets isolated from whole blood from various age groups to evaluate the role of CD63 exosomes during aging.