Exosomes, as extracellular organelles, are the most well-known vesicles among microvesicles. They are released from almost all body fluids as well as cells. Exosomes, which have gained popularity in biology studies with their nano-level dimensions (30-200nm) and membrane structures, are primarily used in drug encapsulation, regeneration, and cell-free cellular therapy. The fact that exosomes, which also function as biomarkers, have an ever-expanding area of use has led researchers to search for the isolation and characterization of this vesicle. In this study, the isolation of exosomes of the DLD-1 colorectal cancer line was carried out by ultracentrifugation method and characterized biophysically and biochemically by SEM, NTA, and flow cytometry, and the proliferative effect of these exosomes on HT29 cells, one of the other colorectal cancer lines, was investigated. As a result, it was proved that the isolated vesicles were morphological and size-appropriate to the literature and expressed CD63 and CD81 proteins. It was observed that DLD-1 exosomes increased proliferation on the HT29 cell line in a dose-dependent manner. Within the framework of the findings, this study could be an excellent reference for exosome isolation and characterization.