The DOI matches its Crossref record; 20 of the 20 references it cites were found in a registry or an index.
Isolation and Characterization of RNA-Containing Exosomes
Open accessAbstract
The field of exosome research is rapidly expanding, with a dramatic increase in publications in recent years. These small vesicles (30-100 nm) of endocytic origin were first proposed to function as a way for reticulocytes to eradicate the transferrin receptor while maturing into erythrocytes, and were later named exosomes. Exosomes are formed by inward budding of late endosomes, producing multivesicular bodies (MVBs), and are released into the environment by fusion of the MVBs with the plasma membrane. Since the first discovery of exosomes, a wide range of cells have been shown to release these vesicles. Exosomes have also been detected in several biological fluids, including plasma, nasal lavage fluid, saliva and breast milk. Furthermore, it has been demonstrated that the content and function of exosomes depends on the originating cell and the conditions under which they are produced. A variety of functions have been demonstrated for exosomes, such as induction of tolerance against allergen, eradication of established tumors in mice, inhibition and activation of natural killer cells, promotion of differentiation into T regulatory cells, stimulation of T cell proliferation and induction of T cell apoptosis. Year 2007 we demonstrated that exosomes released from mast cells contain messenger RNA (mRNA) and microRNA (miRNA), and that the RNA can be shuttled from one cell to another via exosomes. In the recipient cells, the mRNA shuttled by exosomes was shown to be translated into protein, suggesting a regulatory function of the transferred RNA. Further, we have also shown that exosomes derived from cells grown under oxidative stress can induce tolerance against further stress in recipient cells and thus suggest a biological function of the exosomal shuttle RNA. Cell culture media and biological fluids contain a mixture of vesicles and shed fragments. A high quality isolation method for exosomes, followed by characterization and identification of the exosomes and their content, is therefore crucial to distinguish exosomes from other vesicles and particles. Here, we present a method for the isolation of exosomes from both cell culture medium and body fluids. This isolation method is based on repeated centrifugation and filtration steps, followed by a final ultracentrifugation step in which the exosomes are pelleted. Important methods to identify the exosomes and characterize the exosomal morphology and protein content are highlighted, including electron microscopy, flow cytometry and Western blot. The purification of the total exosomal RNA is based on spin column chromatography and the exosomal RNA yield and size distribution is analyzed using a Bioanalyzer.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The cited source says what the claim attributes to it.
In the citing paper
“TSG101 and Alix are two other proteins sometimes used as exosome markers [38, 46, 47].”
From Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies (2013) · cites this paper as [46]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
Valadi, H., Ekström, K., Bossios, A., Sjöstrand, M., Lee, J. J., & Lötvall, J. O. (2007) · Nature Cell Biology · doi:10.1038/ncb1596 · Cited by 12,834
Self-citationB lymphocytes secrete antigen-presenting vesicles.
Raposo, G., Nijman, H. W., Stoorvogel, W., Liejendekker, R., Harding, C. V., Melief, C. J., & Geuze, H. J. (1996) · The Journal of experimental medicine · doi:10.1084/jem.183.3.1161 · Cited by 3,546
Zitvogel, L., Régnault, A., Lozier, A., Wolfers, J., Flament, C., Tenza, D., Ricciardi‐Castagnoli, P., Raposo, G., & Amigorena, S. (1998) · Nature Medicine · doi:10.1038/nm0598-594 · Cited by 2,266
Pan, B., & Johnstone, R. M. (1983) · Cell · doi:10.1016/0092-8674(83)90040-5 · Cited by 2,042
Exosomal-like vesicles are present in human blood plasma
Caby, M.-P., Lankar, D., Vincendeau-Scherrer, C., Raposo, G., & Bonnerot, C. (2005) · International Immunology · doi:10.1093/intimm/dxh267 · Cited by 1,348
Exosomes with Immune Modulatory Features Are Present in Human Breast Milk
Admyre, C., Johansson, S. M., Qazi, K. R., Filén, J., Lahesmaa, R., Norman, M., Neve, E. P. A., Scheynius, A., & Gabrielsson, S. (2007) · The Journal of Immunology · doi:10.4049/jimmunol.179.3.1969 · Cited by 1,222
Human saliva, plasma and breast milk exosomes contain RNA: Uptake by macrophages
Lässer, C., Seyed Alikhani, V., Ekström, K., Eldh, M., Torregrosa Paredes, P., Bossios, A., Sjöstrand, M., Gabrielsson, S., Lötvall, J., & Valadi, H. (2011) · Journal of Translational Medicine · doi:10.1186/1479-5876-9-9 · Cited by 899
Self-citationExosomes: immune properties and potential clinical implementations
Chaput, N., & Théry, C. (2010) · Seminars in Immunopathology · doi:10.1007/s00281-010-0233-9 · Cited by 590
Murine Mammary Carcinoma Exosomes Promote Tumor Growth by Suppression of NK Cell Function
Liu, C., Yu, S., Zinn, K. R., Wang, J., Zhang, L., Jia, Y., Kappes, J. C., Barnes, S., Kimberly, R. P., Grizzle, W. E., et al. (2006) · The Journal of Immunology · doi:10.4049/jimmunol.176.3.1375 · Cited by 499
Szajnik, M. E., Czystowska, M., Szczepański, M. J., Mandapathil, M., & Whiteside, T. L. (2010) · PLoS ONE · doi:10.1371/journal.pone.0011469 · Cited by 478
Flagged first, then by citations
Cited by
Sorted by citations
Biological properties of extracellular vesicles and their physiological functions
Yáñez‐Mó, M., Siljander, P. R. ‐., Andreu, Z., Bedina Zavec, A., Borràs, F. E., Buzas, E. I., Buzas, K., Casal, E., Cappello, F., Carvalho, J., et al. (2015) · Journal of Extracellular Vesicles · doi:10.3402/jev.v4.27066 · Cited by 5,926
Self-citationWitwer, K. W., Buzás, E. I., Bemis, L., Bora, A., Lässer, C., Lötvall, J. O., Hoen, E. N. M. N., Piper, M. G., Sivaraman, S., Skog, J., et al. (2013) · Journal of Extracellular Vesicles · doi:10.3402/jev.v2i0.20360 · Cited by 2,513
Self-citationProgress in Exosome Isolation Techniques
Li, P., Kaslan, M., Lee, S. H., Yao, J. D., & Gao, Z. (2017) · Theranostics · doi:10.7150/thno.18133 · Cited by 2,019
Vlassov, A. V., Magdaleno, S. M., Setterquist, R. A., & Conrad, R. (2012) · Biochimica et Biophysica Acta (BBA) - General Subjects · doi:10.1016/j.bbagen.2012.03.017 · Cited by 1,979
Zhang, H., Freitas, D. P., Kim, H. S., Fabijanic, K. I., Li, Z., Chen, H., Mark, M. T., Molina, H., Benito‐Martín, A., Bojmar, L., et al. (2018) · Nature Cell Biology · doi:10.1038/s41556-018-0040-4 · Cited by 1,666
Metadata
- Authors
- Lässer, C., Eldh, M., & Lötvall, J.
- Affiliations
- University of Gothenburg (SE)
- Venue
- Journal of Visualized Experiments
- Publisher
- MyJove Corporation
- Published
- 2012-01-09
- Issue
- 59
- Pages
- e3037-e3037
- ISSN
- 1940-087X (ISSN-L)
- DOI
- 10.3791/3037
- PMC
- PMC3369768
- OpenAlex
- W2011537287
- Cited by
- 509
- References
- 20
- Citations by year
- 2026: 7 · 2025: 21 · 2024: 36 · 2023: 35 · 2022: 38 · 2021: 47 · 2020: 59 · 2019: 39 · 2018: 47 · 2017: 39 · 2016: 32 · 2015: 39 · 2014: 38 · 2013: 22 · 2012: 10
- FWCI
- 14.02
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- cc-by-nc-nd
- Locations
- Journal of Visualized Experiments (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- Vetenskapsrådet
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- exosomes, exosomal RNA, exosome isolation, ultracentrifugation, multivesicular bodies, exosome characterization, exosomal microRNAs, mRNA transfer, mast cells, oxidative stress, vesicle isolation
- MeSH terms
- Animals, Filtration, Flow Cytometry, Humans, Microscopy, Electron, RNA, Ultracentrifugation, Blotting, Western, Mice, Exosomes
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
