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Residual matrix from different separation techniques impacts exosome biological activity
- Source :
- Scientific Reports
- Publication Year :
- 2016
- Publisher :
- Springer Science and Business Media LLC, 2016.
-
Abstract
- Exosomes are gaining a prominent role in research due to their intriguing biology and several therapeutic opportunities. However, their accurate purification from body fluids and detailed physicochemical characterization remain open issues. We isolated exosomes from serum of patients with Multiple Myeloma by four of the most popular purification methods and assessed the presence of residual contaminants in the preparations through an ad hoc combination of biochemical and biophysical techniques - including Western Blot, colloidal nanoplasmonics, atomic force microscopy (AFM) and scanning helium ion microscopy (HIM). The preparations obtained by iodixanol and sucrose gradients were highly pure. To the contrary, those achieved with limited processing (serial centrifugation or one step precipitation kit) resulted contaminated by a residual matrix, embedding the exosomes. The contaminated preparations showed lower ability to induce NfkB nuclear translocation in endothelial cells with respect to the pure ones, probably because the matrix prevents the interaction and fusion of the exosomes with the cell membrane. These findings suggest that exosome preparation purity must be carefully assessed since it may interfere with exosome biological activity. Contaminants can be reliably probed only by an integrated characterization approach aimed at both the molecular and the colloidal length scales.
- Subjects :
- 0301 basic medicine
Sucrose
Pathology
medicine.medical_specialty
Cell Fractionation
Cell Nucleus
Centrifugation, Density Gradient
Exosomes
Humans
Microscopy, Atomic Force
Multiple Myeloma
NF-kappa B
Protein Transport
Triiodobenzoic Acids
Multidisciplinary
Centrifugation
Biology
Matrix (biology)
Exosome
Article
Cell membrane
03 medical and health sciences
0302 clinical medicine
Western blot
medicine
Multidisciplinary, EXTRACELLULAR VESICLES
FORCE SPECTROSCOPY
HUMAN PLASMA
HUMAN-SALIVA
CANCER
MICROVESICLES
DISEASES
Microscopy
medicine.diagnostic_test
Force spectroscopy
Atomic Force
Biological activity
Microvesicles
Density Gradient
030104 developmental biology
medicine.anatomical_structure
030220 oncology & carcinogenesis
Biophysics
Subjects
Details
- ISSN :
- 20452322
- Volume :
- 6
- Database :
- OpenAIRE
- Journal :
- Scientific Reports
- Accession number :
- edsair.doi.dedup.....f96ca5dff00b1e4d557cb73ca5337c5f