Characterization of extracellular vesicles produced by HEK293 cell line

Author

Rabadán Díaz, Elisabeth 

Abstract

The study of exosomes is an active area of research. Continuous technological and experimental advancements add to our understanding of its biological processes and improve our capacity to use it as a diagnostic and therapeutic tool. Thereby, the aim of this work is to leave the door open to new and more standardized procedures for exosomes characterization. This work is the result of several months experimenting with exosomes in the laboratory. It covers from the biological and therapeutic functions and applications of exosomes for cancer, to their role as drug delivery systems. And for this, the marking of exosomes from HEK293 cell cultures has been studied as well as fluorescence has been demonstrated as a method to characterize them. From HEK293 cell cultures, exosomes were produced, in which fluorescence was applied. Selective labeling of these exosomes was achieved with specific markers on HEK293. By means of bicistronic plasmid cell lines obtained from the laboratory, it was possible to express the CD63 protein, a protein from the tetraspanin family, the main one which was fused with GFP or mCherry to accomplish its fluorescence. Protein expression was evaluated by fluorescence microscopy, flow cytometry, NTA and nanodrop.
The search for characteristics, properties, and functions of exosomes; their active role in labeling them, their potential diagnostic utility and their therapeutic purposes as drug delivery systems have been key to carrying out this work.

Director

Fornaguera i Puigvert, Cristina
Lecina Veciana, Martí

Degree

IQS SE - Undergraduate Program in Pharmacy

Date

2022-09-05