Isolamento e caracterização de microvesículas derivadas de gliomas

Abstract

Background/Aim: Glioblastoma multiforme is the deadliest tumor of central-nervous-system, consisting mainly of tumor and immune cells. T-regulatory cells can modulate to a pro-tumor microenvironment. Microvesicles (MVs), among them exosomes, are responsible of cell-to-cell interaction and are involved in the modulation of tumor-microenvironment. We describe the isolation and characterization of MVs in an in vitro glioma model. Materials and Methods: We performed 4 tests to determine best conditions to isolation of MVs (size and protein content). After determine the best condition, MVs were isolated through ultracentrifugation, characterized by size (ZetaSizer/Nanosight) and protein content. Results: Room temperature thaw and additional centrifugation (test 3) was the test that has the best results for size and protein content. BCA is the better technique to quantify protein concentration of MVs. Nanosight analysis demonstrates minor size variations. MVs show a size increase after five days at 4°C. The treatment with MVs decreases viability of C6 cells in vitro. Conclusion: Successful isolation/characterization of MVs.