% IMPORTANT: The following is UTF-8 encoded. This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.
@ARTICLE{Bordas:157418,
author = {M. Bordas$^*$ and G. Genard$^*$ and S. Ohl$^*$ and M.
Nessling$^*$ and K. Richter$^*$ and T. Roider and S.
Dietrich and K. K. Maaß$^*$ and M. Seiffert$^*$},
title = {{O}ptimized {P}rotocol for {I}solation of {S}mall
{E}xtracellular {V}esicles from {H}uman and {M}urine
{L}ymphoid {T}issues.},
journal = {International journal of molecular sciences},
volume = {21},
number = {15},
issn = {1422-0067},
address = {Basel},
publisher = {Molecular Diversity Preservation International},
reportid = {DKFZ-2020-01613},
pages = {E5586},
year = {2020},
note = {#EA:B060#LA:B060#},
abstract = {Small extracellular vesicles (sEVs) are nanoparticles
responsible for cell-to-cell communication released by
healthy and cancer cells. Different roles have been
described for sEVs in physiological and pathological
contexts, including acceleration of tissue regeneration,
modulation of tumor microenvironment, or premetastatic niche
formation, and they are discussed as promising biomarkers
for diagnosis and prognosis in body fluids. Although efforts
have been made to standardize techniques for isolation and
characterization of sEVs, current protocols often result in
co-isolation of soluble protein or lipid complexes and of
other extracellular vesicles. The risk of contaminated
preparations is particularly high when isolating sEVs from
tissues. As a consequence, the interpretation of data aiming
at understanding the functional role of sEVs remains
challenging and inconsistent. Here, we report an optimized
protocol for isolation of sEVs from human and murine
lymphoid tissues. sEVs from freshly resected human lymph
nodes and murine spleens were isolated comparing two
different approaches-(1) ultracentrifugation on a sucrose
density cushion and (2) combined ultracentrifugation with
size-exclusion chromatography. The purity of sEV
preparations was analyzed using state-of-the-art techniques,
including immunoblots, nanoparticle tracking analysis, and
electron microscopy. Our results clearly demonstrate the
superiority of size-exclusion chromatography, which resulted
in a higher yield and purity of sEVs, and we show that their
functionality alters significantly between the two isolation
protocols.},
cin = {B060 / E041 / W230},
ddc = {540},
cid = {I:(DE-He78)B060-20160331 / I:(DE-He78)E041-20160331 /
I:(DE-He78)W230-20160331},
pnm = {312 - Functional and structural genomics (POF3-312)},
pid = {G:(DE-HGF)POF3-312},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:32759826},
doi = {10.3390/ijms21155586},
url = {https://inrepo02.dkfz.de/record/157418},
}