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@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},
}