Isolation and characterization of exosomes derived from supernatant of rabbit kidney tubular cells culture

Abstract

Introduction: Different vesicles are created by the cells into extra-cellular space, of which researchers mostly considered exosomes. In fact, the vesicles are secreted by each cell type and are found in the body fluids like urine, blood and breast milk and consist of a certain ingredient of proteins, RNA, lipids and DNA. Objectives: The present study attempted to examine the exosomes derived from tubular cells of the rabbit kidney. Materials and Methods: Exosomes have been derived from rabbit kidney tubular cell line (RK13Cs) supernatant by ultracentrifugation centrifugation treatment techniques. In the next stage, RK13Cs-Exo has been validated by the dimension, morphology and certain bio-markers through the transmission electron microscopy (TEM) and scanning electron microscopy (SEM), Zetasizer nano-analysis, atomic force microscopy (AFM) and western blotting. Results: The morphology of RK13Cs-Exo under TEM and SEM revealed that they have been vesicles shaped like a sphere ~150 nm. Zetasizer nano also indicated that RK13Cs-Exo size has been nearly 50 to 100 nm. Atomic force microscopy image of RK13Cs-Exo showed distinct spherical particles in size range between 50 and 100 nm, corresponding to the TEM. According to the western blotting outputs, exosome markers CD63 and CD9 have been expressed in the RK13Cs-Exo. Conclusion: Tubular cells of the rabbit kidney synthesize large amounts of exosomes under critical environmental conditions, which can extract the produced exosomes. According to the characterization of derived exosomes, they are ideal vehicles for drug delivery.

Keywords

• kidney tubular cells
• exosome
• tem
• western blot