Optimization of blood handling for plasma extracellular vesicle isolation
Source abstract
Abstract
Extracellular vesicles (EVs) are emerging targets for biomarker discovery. However, suitable clinical collection and storage methods for blood plasma-derived EVs have not been fully described. We investigated the long- and short-term effects of temperature and time on the plasma-derived EV concentration, size, protein, and RNA content. EVs isolated from freshly collected human blood stored for various periods of time at either room temperature (RT) or 4°C were analyzed for concentration, size, and protein content. In addition, EVs isolated from plasma stored at −80°C were compared to frozen EV pellets. We found that the number, size, protein, and RNA content of EVs were similar between blood samples stored at RT for 24h compared to samples processed shortly after collection or stored at 4°C vs. −80°C. It is therefore feasible to isolate and analyze comparable, stable EVs from blood plasma stored for varying periods of time and at various temperatures.
Source: openaire · Quality: candidate unverified · Source record ↗
Research context
OpenAlex topics: Extracellular vesicles in disease · Platelet Disorders and Treatments · Complement system in diseases
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
References (38)
Show source-provided references
1. Ale Ebrahim S, Ashtari A, Zamani Pedram M, Ale Ebrahim N, Sanati-Nezhad A (2020) Publication trends in exosomes nanoparticles for cancer detection. Int J Nanomedicine 15:4453–4470. https://doi.org/10.2147/IJN.S24721010.2147/IJN.S247210 ↗
2. Anfossi S, Babayan A, Pantel K, Calin GA (2018) Clinical utility of circulating non-coding RNAs - an update. Nat Rev Clin Oncol 15(9):541–563. https://doi.org/10.1038/s41571-018-0035-x10.1038/s41571-018-0035-x ↗
3. Arenaccio C, Federico M (2017) The multifaceted functions of exosomes in health and disease: an overview. Adv Exp Med Biol 998:3–19. https://doi.org/10.1007/978-981-10-4397-0_110.1007/978-981-10-4397-0_1 ↗
4. Baek R, Sondergaard EK, Varming K, Jorgensen MM (2016) The impact of various preanalytical treatments on the phenotype of small extracellular vesicles in blood analyzed by protein microarray. J Immunol Methods 438:11–20. https://doi.org/10.1016/j.jim.2016.08.00710.1016/j.jim.2016.08.007 ↗
5. Baranyai T, Herczeg K, Onodi Z, Voszka I, Modos K, Marton N, Nagy G, Mager I, Wood MJ, El Andaloussi S, Palinkas Z, Kumar V, Nagy P, Kittel A, Buzas EI, Ferdinandy P, Giricz Z (2015) Isolation of exosomes from blood plasma: qualitative and quantitative comparison of ultracentrifugation and size exclusion chromatography methods. PLoS One 10(12):e0145686. https://doi.org/10.1371/journal.pone.014568610.1371/journal.pone.0145686 ↗
6. Barilani M, Peli V, Cherubini A, Dossena M, Dolo V, Lazzari L (2019) NG2 as an identity and quality marker of mesenchymal stem cell extracellular vesicles. Cells 8(12). https://doi.org/10.3390/cells812152410.3390/cells8121524 ↗
7. Bravo V, Rosero S, Ricordi C, Pastori RL (2007) Instability of miRNA and cDNAs derivatives in RNA preparations. Biochem Biophys Res Commun 353(4):1052–1055. https://doi.org/10.1016/j.bbrc.2006.12.13510.1016/j.bbrc.2006.12.135 ↗
8. Cabel L, Proudhon C, Gortais H, Loirat D, Coussy F, Pierga JY, Bidard FC (2017) Circulating tumor cells: clinical validity and utility. Int J Clin Oncol 22(3):421–430. https://doi.org/10.1007/s10147-017-1105-210.1007/s10147-017-1105-2 ↗
9. Chandler WL (2013) Microparticle counts in platelet-rich and platelet-free plasma, effect of centrifugation and sample-processing protocols. Blood Coagul Fibrinolysis 24(2):125–132. https://doi.org/10.1097/MBC.0b013e32835a082410.1097/MBC.0b013e32835a0824 ↗
10. Cheng HH, Yi HS, Kim Y, Kroh EM, Chien JW, Eaton KD, Goodman MT, Tait JF, Tewari M, Pritchard CC (2013) Plasma processing conditions substantially influence circulating microRNA biomarker levels. PLoS One 8(6):e64795. https://doi.org/10.1371/journal.pone.006479510.1371/journal.pone.0064795 ↗
11. Clayton A, Boilard E, Buzas EI, Cheng L, Falcon-Perez JM, Gardiner C, Gustafson D, Gualerzi A, Hendrix A, Hoffman A, Jones J, Lasser C, Lawson C, Lenassi M, Nazarenko I, O’Driscoll L, Pink R, Siljander PR, Soekmadji C, Wauben M, Welsh JA, Witwer K, Zheng L, Nieuwland R (2019) Considerations towards a roadmap for collection, handling and storage of blood extracellular vesicles. J Extracell Vesicles 8(1):1647027. https://doi.org/10.1080/20013078.2019.164702710.1080/20013078.2019.1647027 ↗
12. Cohen JD, Li L, Wang Y, Thoburn C, Afsari B, Danilova L, Douville C, Javed AA, Wong F, Mattox A, Hruban RH, Wolfgang CL, Goggins MG, Dal Molin M, Wang TL, Roden R, Klein AP, Ptak J, Dobbyn L, Schaefer J, Silliman N, Popoli M, Vogelstein JT, Browne JD, Schoen RE, Brand RE, Tie J, Gibbs P, Wong HL, Mansfield AS, Jen J, Hanash SM, Falconi M, Allen PJ, Zhou S, Bettegowda C, Diaz LA Jr, Tomasetti C, Kinzler KW, Vogelstein B, Lennon AM, Papadopoulos N (2018) Detection and localization of surgically resectable cancers with a multi-analyte blood test. Science 359(6378):926–930. https://doi.org/10.1126/science.aar324710.1126/science.aar3247 ↗
13. de Necochea-Campion R, Gonda A, Kabagwira J, Mirshahidi S, Cao H, Reeves ME, Wall NR (2018) A practical approach to extracellular vesicle characterization among similar biological samples. Biomed Phys Eng Express 4:1–810.1088/2057-1976/aad6d8 ↗
14. Doyle LM, Wang MZ (2019) Overview of extracellular vesicles, their origin, composition, purpose, and methods for exosome isolation and analysis. Cells 8(7). https://doi.org/10.3390/cells807072710.3390/cells8070727 ↗
15. Fendl B, Weiss R, Fischer MB, Spittler A, Weber V (2016) Characterization of extracellular vesicles in whole blood: influence of pre-analytical parameters and visualization of vesicle-cell interactions using imaging flow cytometry. Biochem Biophys Res Commun 478(1):168–173. https://doi.org/10.1016/j.bbrc.2016.07.07310.1016/j.bbrc.2016.07.073 ↗
16. Ferguson Bennit HR, Gonda A, McMullen JRW, Kabagwira J, Wall NR (2019) Peripheral blood cell interactions of cancer-derived exosomes affect immune function. Cancer Microenviron 12(1):29–35. https://doi.org/10.1007/s12307-018-0209-110.1007/s12307-018-0209-1 ↗
17. Gonda A, Kabagwira J, Senthil GN, Ferguson Bennit HR, Neidigh JW, Khan S, Wall NR (2018) Exosomal survivin facilitates vesicle internalization. Oncotarget 9(79):34919–34934. https://doi.org/10.18632/oncotarget.2618210.18632/oncotarget.26182 ↗
18. Gonda A, Kabagwira J, Senthil GN, Wall NR (2019) Internalization of exosomes through receptor-mediated endocytosis. Mol Cancer Res 17(2):337–347. https://doi.org/10.1158/1541-7786.MCR-18-089110.1158/1541-7786.MCR-18-0891 ↗
19. Huang T, Deng CX (2019) Current progresses of exosomes as cancer diagnostic and prognostic biomarkers. Int J Biol Sci 15(1):1–11. https://doi.org/10.7150/ijbs.2779610.7150/ijbs.27796 ↗
20. Li W, Li C, Zhou T, Liu X, Liu X, Li X, Chen D (2017) Role of exosomal proteins in cancer diagnosis. Mol Cancer 16(1):145. https://doi.org/10.1186/s12943-017-0706-810.1186/s12943-017-0706-8 ↗
21. Lotvall J, Hill AF, Hochberg F, Buzas EI, Di Vizio D, Gardiner C, Gho YS, Kurochkin IV, Mathivanan S, Quesenberry P, Sahoo S, Tahara H, Wauben MH, Witwer KW, Thery C (2014) Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles. J Extracell Vesicles 3:26913. https://doi.org/10.3402/jev.v3.2691310.3402/jev.v3.26913 ↗
22. Manne U, Jadhav T, Putcha BK, Samuel T, Soni S, Shanmugam C, Suswam EA (2016) Molecular biomarkers of colorectal cancer and cancer disparities: current status and perspective. Curr Colorectal Cancer Rep 12(6):332–344. https://doi.org/10.1007/s11888-016-0338-110.1007/s11888-016-0338-1 ↗
23. Maroto R, Zhao Y, Jamaluddin M, Popov VL, Wang H, Kalubowilage M, Zhang Y, Luisi J, Sun H, Culbertson CT, Bossmann SH, Motamedi M, Brasier AR (2017) Effects of storage temperature on airway exosome integrity for diagnostic and functional analyses. J Extracell Vesicles 6(1):1359478. https://doi.org/10.1080/20013078.2017.135947810.1080/20013078.2017.1359478 ↗
24. Po JW, Roohullah A, Lynch D, DeFazio A, Harrison M, Harnett PR, Kennedy C, de Souza P, Becker TM (2018) Improved ovarian cancer EMT-CTC isolation by immunomagnetic targeting of epithelial EpCAM and mesenchymal N-cadherin. J Circ Biomark 7:1849454418782617. https://doi.org/10.1177/184945441878261710.1177/1849454418782617 ↗
25. Rossi G, Ignatiadis M (2019) Promises and pitfalls of using liquid biopsy for precision medicine. Cancer Res 79(11):2798–2804. https://doi.org/10.1158/0008-5472.CAN-18-340210.1158/0008-5472.CAN-18-3402 ↗
26. Sun Z, Shi K, Yang S, Liu J, Zhou Q, Wang G, Song J, Li Z, Zhang Z, Yuan W (2018) Effect of exosomal miRNA on cancer biology and clinical applications. Mol Cancer 17(1):147. https://doi.org/10.1186/s12943-018-0897-710.1186/s12943-018-0897-7 ↗
27. Taylor DD, Gercel-Taylor C (2013) The origin, function, and diagnostic potential of RNA within extracellular vesicles present in human biological fluids. Front Genet 4:142. https://doi.org/10.3389/fgene.2013.0014210.3389/fgene.2013.00142 ↗
28. Thery C, Amigorena S, Raposo G, Clayton A (2006) Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol Chapter 3(Unit 3):22. https://doi.org/10.1002/0471143030.cb0322s3010.1002/0471143030.cb0322s30 ↗
29. Thery C, Witwer KW, Aikawa E, Alcaraz MJ, Anderson JD, Andriantsitohaina R, Antoniou A, Arab T, Archer F, Atkin-Smith GK, Ayre DC, Bach JM, Bachurski D, Baharvand H, Balaj L, Baldacchino S, Bauer NN, Baxter AA, Bebawy M, Beckham C, Bedina Zavec A, Benmoussa A, Berardi AC, Bergese P, Bielska E, Blenkiron C, Bobis-Wozowicz S, Boilard E, Boireau W, Bongiovanni A, Borras FE, Bosch S, Boulanger CM, Breakefield X, Breglio AM, Brennan MA, Brigstock DR, Brisson A, Broekman ML, Bromberg JF, Bryl-Gorecka P, Buch S, Buck AH, Burger D, Busatto S, Buschmann D, Bussolati B, Buzas EI, Byrd JB, Camussi G, Carter DR, Caruso S, Chamley LW, Chang YT, Chen C, Chen S, Cheng L, Chin AR, Clayton A, Clerici SP, Cocks A, Cocucci E, Coffey RJ, Cordeiro-da-Silva A, Couch Y, Coumans FA, Coyle B, Crescitelli R, Criado MF, D’Souza-Schorey C, Das S, Datta Chaudhuri A, de Candia P, De Santana EF, De Wever O, Del Portillo HA, Demaret T, Deville S, Devitt A, Dhondt B, Di Vizio D, Dieterich LC, Dolo V, Dominguez Rubio AP, Dominici M, Dourado MR, Driedonks TA, Duarte FV, Duncan HM, Eichenberger RM, Ekstrom K, El Andaloussi S, Elie-Caille C, Erdbrugger U, Falcon-Perez JM, Fatima F, Fish JE, Flores-Bellver M, Forsonits A, Frelet-Barrand A, Fricke F, Fuhrmann G, Gabrielsson S, Gamez-Valero A, Gardiner C, Gartner K, Gaudin R, Gho YS, Giebel B, Gilbert C, Gimona M, Giusti I, Goberdhan DC, Gorgens A, Gorski SM, Greening DW, Gross JC, Gualerzi A, Gupta GN, Gustafson D, Handberg A, Haraszti RA, Harrison P, Hegyesi H, Hendrix A, Hill AF, Hochberg FH, Hoffmann KF, Holder B, Holthofer H, Hosseinkhani B, Hu G, Huang Y, Huber V, Hunt S, Ibrahim AG, Ikezu T, Inal JM, Isin M, Ivanova A, Jackson HK, Jacobsen S, Jay SM, Jayachandran M, Jenster G, Jiang L, Johnson SM, Jones JC, Jong A, Jovanovic-Talisman T, Jung S, Kalluri R, Kano SI, Kaur S, Kawamura Y, Keller ET, Khamari D, Khomyakova E, Khvorova A, Kierulf P, Kim KP, Kislinger T, Klingeborn M, Klinke DJ 2nd, Kornek M, Kosanovic MM, Kovacs AF, Kramer-Albers EM, Krasemann S, Krause M, Kurochkin IV, Kusuma GD, Kuypers S, Laitinen S, Langevin SM, Languino LR, Lannigan J, Lasser C, Laurent LC, Lavieu G, Lazaro-Ibanez E, Le Lay S, Lee MS, Lee YXF, Lemos DS, Lenassi M, Leszczynska A, Li IT, Liao K, Libregts SF, Ligeti E, Lim R, Lim SK, Line A, Linnemannstons K, Llorente A, Lombard CA, Lorenowicz MJ, Lorincz AM, Lotvall J, Lovett J, Lowry MC, Loyer X, Lu Q, Lukomska B, Lunavat TR, Maas SL, Malhi H, Marcilla A, Mariani J, Mariscal J, Martens-Uzunova ES, Martin-Jaular L, Martinez MC, Martins VR, Mathieu M, Mathivanan S, Maugeri M, LK MG, MJ MV, Meckes DG Jr, Meehan KL, Mertens I, Minciacchi VR, Moller A, Moller Jorgensen M, Morales-Kastresana A, Morhayim J, Mullier F, Muraca M, Musante L, Mussack V, Muth DC, Myburgh KH, Najrana T, Nawaz M, Nazarenko I, Nejsum P, Neri C, Neri T, Nieuwland R, Nimrichter L, Nolan JP, Nolte-‘t Hoen EN, Noren Hooten N, O’Driscoll L, O’Grady T, O’Loghlen A, Ochiya T, Olivier M, Ortiz A, Ortiz LA, Osteikoetxea X, Ostergaard O, Ostrowski M, Park J, Pegtel DM, Peinado H, Perut F, Pfaffl MW, Phinney DG, Pieters BC, Pink RC, Pisetsky DS, Pogge von Strandmann E, Polakovicova I, Poon IK, Powell BH, Prada I, Pulliam L, Quesenberry P, Radeghieri A, Raffai RL, Raimondo S, Rak J, Ramirez MI, Raposo G, Rayyan MS, Regev-Rudzki N, Ricklefs FL, Robbins PD, Roberts DD, Rodrigues SC, Rohde E, Rome S, Rouschop KM, Rughetti A, Russell AE, Saa P, Sahoo S, Salas-Huenuleo E, Sanchez C, Saugstad JA, Saul MJ, Schiffelers RM, Schneider R, Schoyen TH, Scott A, Shahaj E, Sharma S, Shatnyeva O, Shekari F, Shelke GV, Shetty AK, Shiba K, Siljander PR, Silva AM, Skowronek A, Snyder OL 2nd, Soares RP, Sodar BW, Soekmadji C, Sotillo J, Stahl PD, Stoorvogel W, Stott SL, Strasser EF, Swift S, Tahara H, Tewari M, Timms K, Tiwari S, Tixeira R, Tkach M, Toh WS, Tomasini R, Torrecilhas AC, Tosar JP, Toxavidis V, Urbanelli L, Vader P, van Balkom BW, van der Grein SG, Van Deun J, van Herwijnen MJ, Van Keuren-Jensen K, van Niel G, van Royen ME, van Wijnen AJ, Vasconcelos MH, Vechetti IJ Jr, Veit TD, Vella LJ, Velot E, Verweij FJ, Vestad B, Vinas JL, Visnovitz T, Vukman KV, Wahlgren J, Watson DC, Wauben MH, Weaver A, Webber JP, Weber V, Wehman AM, Weiss DJ, Welsh JA, Wendt S, Wheelock AM, Wiener Z, Witte L, Wolfram J, Xagorari A, Xander P, Xu J, Yan X, Yanez-Mo M, Yin H, Yuana Y, Zappulli V, Zarubova J, Zekas V, Zhang JY, Zhao Z, Zheng L, Zheutlin AR, Zickler AM, Zimmermann P, Zivkovic AM, Zocco D, Zuba-Surma EK (2018) Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles 7(1):1535750. https://doi.org/10.1080/20013078.2018.153575010.1080/20013078.2018.1535750 ↗
30. Turay D, Khan S, Diaz Osterman CJ, Curtis MP, Khaira B, Neidigh JW, Mirshahidi S, Casiano CA, Wall NR (2016) Proteomic profiling of serum-derived exosomes from ethnically diverse prostate cancer patients. Cancer Investig 34(1):1–11. https://doi.org/10.3109/07357907.2015.108192110.3109/07357907.2015.1081921 ↗
31. Valenzuela M, Ferguson Bennit H, Gonda A, Diaz Osterman C, Hibma A, Khan S, Wall N (2015) Exosomes secreted from human cancer cell lines contain inhibitors of apoptosis (IAP). Cancer Microenviron 8(2):65–73. 1-9. https://doi.org/10.1007/s12307-015-0167-910.1007/s12307-015-0167-9 ↗
32. Volckmar AL, Sultmann H, Riediger A, Fioretos T, Schirmacher P, Endris V, Stenzinger A, Dietz S (2018) A field guide for cancer diagnostics using cell-free DNA: from principles to practice and clinical applications. Genes Chromosom Cancer 57(3):123–139. https://doi.org/10.1002/gcc.2251710.1002/gcc.22517 ↗
33. Wisgrill L, Lamm C, Hartmann J, Preissing F, Dragosits K, Bee A, Hell L, Thaler J, Ay C, Pabinger I, Berger A, Spittler A (2016) Peripheral blood microvesicles secretion is influenced by storage time, temperature, and anticoagulants. Cytometry A 89(7):663–672. https://doi.org/10.1002/cyto.a.2289210.1002/cyto.a.22892 ↗
34. Witwer KW, Buzas EI, Bemis LT, Bora A, Lasser C, Lotvall J, Nolte-’t Hoen EN, Piper MG, Sivaraman S, Skog J, Thery C, Wauben MH, Hochberg F (2013) Standardization of sample collection, isolation and analysis methods in extracellular vesicle research. J Extracell Vesicles 2. https://doi.org/10.3402/jev.v2i0.2036010.3402/jev.v2i0.20360 ↗
35. Yamashita T, Takahashi Y, Takakura Y (2018) Possibility of exosome-based therapeutics and challenges in production of exosomes eligible for therapeutic application. Biol Pharm Bull 41(6):835–842. https://doi.org/10.1248/bpb.b18-0013310.1248/bpb.b18-00133 ↗
36. Yanez-Mo M, Siljander PR, Andreu Z, Zavec AB, Borras FE, Buzas EI, Buzas K, Casal E, Cappello F, Carvalho J, Colas E, Cordeiro-da Silva A, Fais S, Falcon-Perez JM, Ghobrial IM, Giebel B, Gimona M, Graner M, Gursel I, Gursel M, Heegaard NH, Hendrix A, Kierulf P, Kokubun K, Kosanovic M, Kralj-Iglic V, Kramer-Albers EM, Laitinen S, Lasser C, Lener T, Ligeti E, Line A, Lipps G, Llorente A, Lotvall J, Mancek-Keber M, Marcilla A, Mittelbrunn M, Nazarenko I, Nolte-’t Hoen EN, Nyman TA, O’Driscoll L, Olivan M, Oliveira C, Pallinger E, Del Portillo HA, Reventos J, Rigau M, Rohde E, Sammar M, Sanchez-Madrid F, Santarem N, Schallmoser K, Ostenfeld MS, Stoorvogel W, Stukelj R, Van der Grein SG, Vasconcelos MH, Wauben MH, De Wever O (2015) Biological properties of extracellular vesicles and their physiological functions. J Extracell Vesicles 4:27066. https://doi.org/10.3402/jev.v4.2706610.3402/jev.v4.27066 ↗
37. Yuana Y, Boing AN, Grootemaat AE, van der Pol E, Hau CM, Cizmar P, Buhr E, Sturk A, Nieuwland R (2015) Handling and storage of human body fluids for analysis of extracellular vesicles. J Extracell Vesicles 4:29260. https://doi.org/10.3402/jev.v4.2926010.3402/jev.v4.29260 ↗
38. Zaborowski MP, Balaj L, Breakefield XO, Lai CP (2015) Extracellular vesicles: composition, biological relevance, and methods of study. Bioscience 65(8):783–797. https://doi.org/10.1093/biosci/biv08410.1093/biosci/biv084 ↗