Characterization of Exosomes Isolated from Cell Culture Media

Exosomes are intensively studied as potential drug delivery vehicles. Here we demonstrate that the Litesizer™ 500 can efficiently characterize exosome particle size, and that this criterion can be used to monitor exosome stability in vitro. Zeta potential measurements were also performed, giving potentially useful information about the biological functionality of exosomes.

Exosomes are nanometer-range vesicles originating from the fusion of endosomes with the plasma membrane of eukaryotic cells. They can be isolated from cell culture medium as well as from many extracellular fluids such as blood, urine, saliva, breast milk and cerebrospinal fluid. 

Based on electron microscopy analysis, exosomes are tiny vesicles with an expected size range of 30 to 150 nm. They have a lipid bilayer similar to liposomes but are more heterogeneous in size and have a more complex make-up. They can incorporate proteins (e.g., Tetraspanins), mRNA, miRNA, cholesterol, sphingomyelin and mostly mono-unsaturated or saturated fatty acids. 

While the in vivo physiological relevance of exosomes is still largely unclear, their potential as drug delivery system has long been recognized.  Their many advantages include a long-range cell-targeting action, low toxicity, low immunogenicity, high stability and the capacity to encapsulate proteins, drugs or nucleic acids. As such, they are regarded as one of the best candidates for cancer-targeting therapy. In addition, they are becoming powerful diagnostic tools, as the RNA and protein profile of circulating exosomes can be associated with an increasing range of diseases.

In this report, exosomes isolated from the cell culture medium of human buccal cells were characterized using a Litesizer™ 500.

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