Tangential Flow Filtration for Highly Efficient Concentration of Extracellular Vesicles from Large Volumes of Fluid
by
Sara Busatto 1,2,*,†, George Vilanilam 1,†
, Taylor Ticer 1, Wen-Lang Lin 3, Dennis W. Dickson 3, Shane Shapiro 4, Paolo Bergese 2,5,* and Joy Wolfram 1,6,*
Sara Busatto 1,2,*,†, George Vilanilam 1,†, Taylor Ticer 1, Wen-Lang Lin 3, Dennis W. Dickson 3, Shane Shapiro 4, Paolo Bergese 2,5,* and Joy Wolfram 1,6,*
1
Department of Transplantation Medicine; Department of Physiology and Biomedical Engineering, Mayo Clinic, Jacksonville, FL 32224, USA
2
Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy
3
Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA
4
Department of Orthopedic Surgery, Mayo Clinic, Jacksonville, FL 32224, USA
5
CSGI, Research Center for Colloids and Nanoscience, 50019 Florence, Italy
6
Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030, USA
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Cells 2018, 7(12), 273; https://doi.org/10.3390/cells7120273
Received: 11 October 2018 / Revised: 2 December 2018 / Accepted: 14 December 2018 / Published: 16 December 2018
(This article belongs to the Special Issue Exosomes and Extracellular Vesicles in Health and Disease)
Concentration of extracellular vesicles (EVs) from biological fluids in a scalable and reproducible manner represents a major challenge. This study reports the use of tangential flow filtration (TFF) for the highly efficient isolation of EVs from large volumes of samples. When compared to ultracentrifugation (UC), which is the most widely used method to concentrate EVs, TFF is a more efficient, scalable, and gentler method. Comparative assessment of TFF and UC of conditioned cell culture media revealed that the former concentrates EVs of comparable physicochemical characteristics, but with higher yield, less single macromolecules and aggregates (<15 nm in size), and improved batch-to-batch consistency in half the processing time (1 h). The TFF protocol was then successfully implemented on fluids derived from patient lipoaspirate. EVs from adipose tissue are of high clinical relevance, as they are expected to mirror the regenerative properties of the parent cells.
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Keywords:
exosome; extracellular vesicles; isolation; ultrafiltration; tangential flow filtration; lipoaspirate
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MDPI and ACS Style
Busatto, S.; Vilanilam, G.; Ticer, T.; Lin, W.-L.; Dickson, D.W.; Shapiro, S.; Bergese, P.; Wolfram, J. Tangential Flow Filtration for Highly Efficient Concentration of Extracellular Vesicles from Large Volumes of Fluid. Cells 2018, 7, 273.
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