FREE SHIPPING over $40
OVER 10000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
FASEB journal : official publication of the Federation of American Societies for Experimental Biology2026; 40(2); e71472; doi: 10.1096/fj.202504053R

Comparative Analysis of Extracellular Vesicle Isolation From Equine Serum and Plasma Using Two Isolation Methods With Structural and Proteomic Validation.

Abstract: Extracellular vesicles (EVs) are promising biomarkers and mediators of intercellular communication, but their isolation from equine biofluids remains challenging. This study compared two isolation workflows-size-exclusion chromatography (SEC) and differential ultracentrifugation followed by SEC (UC + SEC)-to evaluate their efficiency, reproducibility, and the proteomic composition of EVs derived from equine serum and plasma. Blood from six healthy horses was processed to obtain platelet-free plasma and serum. EVs were isolated using SEC or UC + SEC and characterized by transmission electron microscopy, nanoparticle tracking analysis, and mass spectrometry-based proteomics with functional enrichment analysis. SEC provided higher reproducibility, greater protein yield, and a simpler workflow than UC + SEC. Serum combined with SEC yielded the most consistent proteomic profiles, with strong detection of typical EV markers and the largest overlap among replicates. Vesicles displayed the expected morphology and size distribution, with a predominant population between 100 and 200 nm. Plasma-derived EVs were enriched in proteins related to translation, chaperone activity, and proteasome function, while serum-derived EVs contained proteins involved in immune processes, cytoskeletal organization, adhesion, and hemostasis. Both the isolation method and biological matrix significantly influenced EV yield and proteome composition. SEC applied to serum provided a reproducible and high-quality EV preparation suitable for biomarker discovery. As this was a methodological comparison in healthy animals, diagnostic test performance metrics (sensitivity, specificity, PPV/NPV) were outside the scope of the study; our goal was to quantify upstream analytical determinants (matrix and isolation workflow) that influence reproducibility and discovery-phase proteomic readouts.
© 2026 The Author(s). The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.
Get Full Text
Publication Date: 2026-01-19 PubMed ID: 41549528PubMed Central: PMC12813514DOI: 10.1096/fj.202504053RGoogle Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Comparative Study

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Extracellular vesicles (EVs) were isolated from horse serum and plasma using two different methods to determine which provides better yield, reproducibility, and protein profiling. The study found that size-exclusion chromatography alone on serum samples yielded the most consistent and high-quality EV preparations.

Background and Purpose

  • Extracellular vesicles (EVs) are small particles secreted by cells that carry proteins and other molecules, playing important roles in cell communication and serving as potential biomarkers for diseases.
  • Isolation of EVs from equine (horse) biofluids like serum and plasma is difficult due to the complex composition of these fluids.
  • This study aimed to compare two common EV isolation workflows—size-exclusion chromatography (SEC) alone and differential ultracentrifugation followed by SEC (UC + SEC)—to evaluate which provides better efficiency, reproducibility, and proteomic profiles in horses.
  • Understanding the best method helps facilitate biomarker discovery in equine medicine by ensuring reliable EV isolation.

Methods

  • Blood samples were collected from six healthy horses.
  • The samples were processed to obtain platelet-free plasma and serum.
  • EVs were isolated using either:
    • Size-exclusion chromatography (SEC) alone
    • Differential ultracentrifugation followed by SEC (UC + SEC)
  • Several characterization techniques were applied:
    • Transmission Electron Microscopy (TEM) to visualize the vesicles’ morphology
    • Nanoparticle tracking analysis (NTA) to measure size distribution and concentration
    • Mass spectrometry-based proteomics with functional enrichment analysis to detail protein composition

Key Findings

  • Isolation Method Efficiency and Reproducibility:
    • SEC alone was more reproducible and simpler. It yielded greater amounts of protein compared to UC + SEC.
    • UC + SEC was less consistent, more complex, and resulted in lower protein yield.
  • Biological Matrix Impact:
    • Serum combined with SEC produced the most consistent proteomic profiles across replicates.
    • Serum EV preparations showed strong detection of typical EV marker proteins and a high overlap of protein identities between samples.
    • Plasma EVs and serum EVs differed in protein composition, reflecting distinct biological functions and processes.
  • EV Morphology and Size:
    • Both isolation methods yielded vesicles exhibiting expected morphology typical for EVs under electron microscopy.
    • Size distributions were predominantly in the 100–200 nm range, consistent across methods and matrices.
  • Proteomic Differences:
    • Plasma-derived EVs were enriched with proteins linked to:
      • Translation machinery
      • Chaperone activity
      • Proteasome functions (protein degradation)
    • Serum-derived EVs contained proteins related to:
      • Immune processes
      • Cytoskeleton organization
      • Cell adhesion
      • Hemostasis (blood clotting)

Conclusions and Implications

  • Both the choice of biofluid (serum vs plasma) and the isolation method significantly impact EV yield and proteomic content.
  • SEC applied to serum samples offers a reproducible, simpler, and higher-quality approach for isolating EVs suitable for downstream proteomic analyses and biomarker discovery in horses.
  • The study focused on methodological comparison rather than diagnostic performance metrics, providing foundational knowledge for future studies that might evaluate clinically relevant EV biomarkers.
  • This optimized isolation workflow can aid research into veterinary diagnostics and improve understanding of equine diseases through EV analysis.

Cite This Article

APA
Milczek-Haduch D, Żmigrodzka M, Kiełbik P, Świderska B, Olędzki J, Witkowska-Piłaszewicz O. (2026). Comparative Analysis of Extracellular Vesicle Isolation From Equine Serum and Plasma Using Two Isolation Methods With Structural and Proteomic Validation. FASEB J, 40(2), e71472. https://doi.org/10.1096/fj.202504053R

Publication

FASEB journal : official publication of the Federation of American Societies for Experimental Biology
ISSN: 1530-6860
NlmUniqueID: 8804484
Country: United States
Language: English
Volume: 40
Issue: 2
Pages: e71472
PII: e71472

Researcher Affiliations

Milczek-Haduch, Dominika
  • Department of Large Animals Diseases and Clinic, Institute of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.
  • Department of Morphological Sciences, Institute of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.
Żmigrodzka, Magdalena
  • Department of Pathology and Veterinary Diagnostic, Institute of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.
Kiełbik, Paula
  • Department of Large Animals Diseases and Clinic, Institute of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.
Świderska, Bianka
  • Mass Spectrometry Laboratory, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
Olędzki, Jacek
  • Mass Spectrometry Laboratory, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
Witkowska-Piłaszewicz, Olga
  • Department of Large Animals Diseases and Clinic, Institute of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.

MeSH Terms

  • Animals
  • Horses / blood
  • Extracellular Vesicles / metabolism
  • Extracellular Vesicles / ultrastructure
  • Proteomics / methods
  • Plasma / metabolism
  • Plasma / chemistry
  • Proteome / metabolism
  • Chromatography, Gel / methods
  • Biomarkers / blood
  • Ultracentrifugation / methods
  • Serum / metabolism
  • Serum / chemistry

Grant Funding

  • 2022/47/D/NZ7/01814 / Narodowe Centrum Nauki (NCN)
  • the Science Development Fund of The Warsaw Univeristy of Life Sciences-SGGW

Conflict of Interest Statement

The authors declare no conflicts of interest.

References

This article includes 74 references
  1. Park J, Park J S, Huang C H. An Integrated Magneto‐Electrochemical Device for the Rapid Profiling of Tumour Extracellular Vesicles From Blood Plasma. Nature Biomedical Engineering 5, no. 7 (2021): 678–689.
    pmc: PMC8437135pubmed: 34183802
  2. Ferguson S, Yang K S, Zelga P. Single‐EV Analysis (sEVA) of Mutated Proteins Allows Detection of Stage 1 Pancreatic Cancer. Science Advances 8, no. 16 (2022): eabm3453.
    pmc: PMC9032977pubmed: 35452280
  3. Chatterjee M, Özdemir S, Fritz C. Plasma Extracellular Vesicle Tau and TDP‐43 as Diagnostic Biomarkers in FTD and ALS. Nature Medicine 30, no. 6 (2024): 1771–1783.
    pmc: PMC11186765pubmed: 38890531
  4. Xiong Y, Lou P, Xu C, Han B, Liu J, Gao J. Emerging Role of Extracellular Vesicles in Veterinary Practice: Novel Opportunities and Potential Challenges. Frontiers in Veterinary Science 11 (2024): 1335107.
    pmc: PMC10850357pubmed: 38332755
  5. Théry C, Witwer K W, Aikawa E. 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. Journal of Extracellular Vesicles 7, no. 1 (2018): 1535750.
    pmc: PMC6322352pubmed: 30637094
  6. Welsh J A, Goberdhan D C I, O'Driscoll L. Minimal Information for Studies of Extracellular Vesicles (MISEV2023): From Basic to Advanced Approaches. Journal of Extracellular Vesicles 13, no. 2 (2024): e12404.
    pmc: PMC10850029pubmed: 38326288
  7. Soukup R, Gerner I, Gültekin S. Characterisation of Extracellular Vesicles From Equine Mesenchymal Stem Cells. International Journal of Molecular Sciences 23, no. 10 (2022): 5858.
    pmc: PMC9145091pubmed: 35628667
  8. Zhang X, Takeuchi T, Takeda A, Mochizuki H, Nagai Y. Comparison of Serum and Plasma as a Source of Blood Extracellular Vesicles: Increased Levels of Platelet‐Derived Particles in Serum Extracellular Vesicle Fractions Alter Content Profiles From Plasma Extracellular Vesicle Fractions. PLoS One 17, no. 6 (2022): e0270634.
    pmc: PMC9231772pubmed: 35749554
  9. Taha H B. Plasma Versus Serum for Extracellular Vesicle (EV) Isolation: A Duel for Reproducibility and Accuracy for CNS‐Originating EVs Biomarker Analysis. Journal of Neuroscience Research 101, no. 11 (2023): 1677–1686.
    pubmed: 37501394
  10. Royo F, Théry C, Falcón‐Pérez J M, Nieuwland R, Witwer K W. Methods for Separation and Characterization of Extracellular Vesicles: Results of a Worldwide Survey Performed by the ISEV Rigor and Standardization Subcommittee. Cells 9, no. 9 (2020): 1955.
    pmc: PMC7563174pubmed: 32854228
  11. Combeaud E, Touitou F, Bret L. Lipoproteins and Cholesterol Transport in Dogs, Cats and Horses: Particular Feature Compared to Humans (Mini Review). Biomedical Journal of Scientific & Technical Research 43, no. 2 (2022): 34479–34487.
  12. Watson T D G, Burns L, Love S, Packard C J, Shepherd J. The Isolation, Characterisation and Quantification of the Equine Plasma Lipoproteins. Equine Veterinary Journal 23, no. 5 (1991): 353–359.
    pubmed: 1959526
  13. Monguió‐Tortajada M, Gálvez‐Montón C, Bayes‐Genis A, Roura S, Borràs F E. Extracellular Vesicle Isolation Methods: Rising Impact of Size‐Exclusion Chromatography. Cellular and Molecular Life Sciences 76, no. 12 (2019): 2369–2382.
    pmc: PMC11105396pubmed: 30891621
  14. Sidhom K, Obi P O, Saleem A. A Review of Exosomal Isolation Methods: Is Size Exclusion Chromatography the Best Option?. International Journal of Molecular Sciences 21, no. 18 (2020): 6466.
    pmc: PMC7556044pubmed: 32899828
  15. Bullone M, Lavoie J P. The Equine Asthma Model of Airway Remodeling: From a Veterinary to a Human Perspective. Cell and Tissue Research 380, no. 2 (2020): 223–236.
    pubmed: 31713728
  16. Couetil L, Cardwell J M, Leguillette R. Equine Asthma: Current Understanding and Future Directions. Frontiers in Veterinary Science 7 (2020): 450.
    pmc: PMC7438831pubmed: 32903600
  17. Benammar A, Derisoud E, Vialard F. The Mare: A Pertinent Model for Human Assisted Reproductive Technologies?. Animals 11, no. 8 (2021): 2304.
    pmc: PMC8388489pubmed: 34438761
  18. Rizzo M, Du Preez N, Ducheyne K D. The Horse as a Natural Model to Study Reproductive Aging‐Induced Aneuploidy and Weakened Centromeric Cohesion in Oocytes. Aging 12, no. 21 (2020): 22220–22232.
    pmc: PMC7695376pubmed: 33139583
  19. Witkowska‐Piłaszewicz O, Malin K, Dąbrowska I, Grzędzicka J, Ostaszewski P, Carter C. Immunology of Physical Exercise: Is an Appropriate Animal Model for Human Athletes?. International Journal of Molecular Sciences 25, no. 10 (2024): 5210.
    pmc: PMC11121269pubmed: 38791248
  20. McIlwraith C W, Frisbie D D, Kawcak C E. The Horse as a Model of Naturally Occurring Osteoarthritis. Bone & Joint Research 1, no. 11 (2012): 297–309.
    pmc: PMC3626203pubmed: 23610661
  21. Ertelt A, Barton A K, Schmitz R R, Gehlen H. Metabolic Syndrome: Is Equine Disease Comparable to What We Know in Humans?. Endocrine Connections 3, no. 3 (2014): R81–R93.
    pmc: PMC4068110pubmed: 24894908
  22. Schüttler D, Bapat A, Kääb S. Animal Models of Atrial Fibrillation. Circulation Research 127, no. 1 (2020): 91–110.
    pubmed: 32716814
  23. Hesselkilde E Z, Carstensen H, Flethøj M. Longitudinal Study of Electrical, Functional and Structural Remodelling in an Equine Model of Atrial Fibrillation. BMC Cardiovascular Disorders 19, no. 1 (2019): 228.
    pmc: PMC6805623pubmed: 31638896
  24. Patterson‐Kane J C, Rich T. Achilles Tendon Injuries in Elite Athletes: Lessons in Pathophysiology From Their Equine Counterparts. ILAR Journal 55, no. 1 (2014): 86–99.
    pubmed: 24936032
  25. Van Deun J, Mestdagh P, Sormunen R. The Impact of Disparate Isolation Methods for Extracellular Vesicles on Downstream RNA Profiling. Journal of Extracellular Vesicles 3, no. 3 (2014).
    doi: 10.3402/jev.v3.24858pmc: PMC4169610pubmed: 25317274google scholar: lookup
  26. Nieuwland R, Siljander P R. A Beginner's Guide to Study Extracellular Vesicles in Human Blood Plasma and Serum. Journal of Extracellular Vesicles 13, no. 1 (2024): e12400.
    pmc: PMC10775135pubmed: 38193375
  27. Ludwig N, Hong C, Ludwig S. Isolation and Analysis of Tumor‐Derived Exosomes. Current Protocols in Immunology 127, no. 1 (2019): e91.
    pmc: PMC6880756pubmed: 31763776
  28. Dlugolecka M, Szymanski J, Zareba L. Characterization of Extracellular Vesicles From Bronchoalveolar Lavage Fluid and Plasma of Patients With Lung Lesions Using Fluorescence Nanoparticle Tracking Analysis. Cells 10, no. 12 (2021): 3473.
    pmc: PMC8699990pubmed: 34943982
  29. Holcar M, Ferdin J, Sitar S. Enrichment of Plasma Extracellular Vesicles for Reliable Quantification of Their Size and Concentration for Biomarker Discovery. Scientific Reports 10, no. 1 (2020): 21346.
    pmc: PMC7721811pubmed: 33288809
  30. Zhang K, Dong C, Chen M. Extracellular Vesicle‐Mediated Delivery of miR‐101 Inhibits Lung Metastasis in Osteosarcoma. Theranostics 10, no. 1 (2020): 411–425.
    pmc: PMC6929625pubmed: 31903129
  31. Zhang Q, Jeppesen D K, Higginbotham J N, Franklin J L, Coffey R J. Comprehensive Isolation of Extracellular Vesicles and Nanoparticles. Nature Protocols 18, no. 5 (2023): 1462–1487.
    pmc: PMC10445291pubmed: 36914899
  32. Hughes C S, Moggridge S, Müller T, Sorensen P H, Morin G B, Krijgsveld J. Single‐Pot, Solid‐Phase‐Enhanced Sample Preparation for Proteomics Experiments. Nature Protocols 14, no. 1 (2019): 68–85.
    pubmed: 30464214
  33. Vallejo M C, Sarkar S, Elliott E C. A Proteomic Meta‐Analysis Refinement of Plasma Extracellular Vesicles. Scientific Data 10, no. 1 (2023): 837.
    pmc: PMC10684639pubmed: 38017024
  34. Mol E A, Goumans M J, Doevendans P A, Sluijter J P G, Vader P. Higher Functionality of Extracellular Vesicles Isolated Using Size‐Exclusion Chromatography Compared to Ultracentrifugation. Nanomedicine: Nanotechnology, Biology, and Medicine 13, no. 6 (2017): 2061–2065.
    pubmed: 28365418
  35. Lobb R J, Becker M, Wen S W. Optimized Exosome Isolation Protocol for Cell Culture Supernatant and Human Plasma. Journal of Extracellular Vesicles 4 (2015): 27031.
    pmc: PMC4507751pubmed: 26194179
  36. Lamparski H G, Metha‐Damani A, Yao J Y. Production and Characterization of Clinical Grade Exosomes Derived From Dendritic Cells. Journal of Immunological Methods 270, no. 2 (2002): 211–226.
    pubmed: 12379326
  37. Langevin S M, Kuhnell D, Orr‐Asman M A. Balancing Yield, Purity and Practicality: A Modified Differential Ultracentrifugation Protocol for Efficient Isolation of Small Extracellular Vesicles From Human Serum. RNA Biology 16, no. 1 (2019): 5–12.
    pmc: PMC6380284pubmed: 30604646
  38. Konoshenko M Y, Lekchnov E A, Vlassov A V, Laktionov P P. Isolation of Extracellular Vesicles: General Methodologies and Latest Trends. BioMed Research International 2018 (2018): 1–27.
    pmc: PMC5831698pubmed: 29662902
  39. Benayas B, Morales J, Egea C, Armisén P, Yáñez‐Mó M. Optimization of Extracellular Vesicle Isolation and Their Separation From Lipoproteins by Size Exclusion Chromatography. Journal of Extracellular Biology 2, no. 7 (2023): e100.
    pmc: PMC11080862pubmed: 38939075
  40. Reshi Q U A, Hasan M M, Dissanayake K, Fazeli A. Isolation of Extracellular Vesicles (EVs) Using Benchtop Size Exclusion Chromatography (SEC) Columns. Methods in Molecular Biology 2273 (2021): 201–206.
    pubmed: 33604855
  41. Yuana Y, Levels J, Grootemaat A, Sturk A, Nieuwland R. Co‐Isolation of Extracellular Vesicles and High‐Density Lipoproteins Using Density Gradient Ultracentrifugation. Journal of Extracellular Vesicles 3 (2014): 3.
    pmc: PMC4090368pubmed: 25018865
  42. Yang Y, Wang Y, Wei S. Extracellular Vesicles Isolated by Size‐Exclusion Chromatography Present Suitability for RNomics Analysis in Plasma. Journal of Translational Medicine 19, no. 1 (2021): 104.
    pmc: PMC7953782pubmed: 33712033
  43. Gámez‐Valero A, Monguió‐Tortajada M, Carreras‐Planella L, Franquesa M, Beyer K, Borràs F E. Size‐Exclusion Chromatography‐Based Isolation Minimally Alters Extracellular Vesicles' Characteristics Compared to Precipitating Agents. Scientific Reports 6, no. 1 (2016): 33641.
    pmc: PMC5027519pubmed: 27640641
  44. Merij L B, da Silva L R, Palhinha L. Density‐Based Lipoprotein Depletion Improves Extracellular Vesicle Isolation and Functional Analysis. Journal of Thrombosis and Haemostasis 22, no. 5 (2024): 1372–1388.
    pubmed: 38278418
  45. Bettio V, Mazzucco E, Antona A. Extracellular Vesicles From Human Plasma for Biomarkers Discovery: Impact of Anticoagulants and Isolation Techniques. PLoS One 18, no. 5 (2023): e0285440.
    pmc: PMC10171685pubmed: 37163560
  46. George J N, Thoi L L, McManus L M, Reimann T A. Isolation of Human Platelet Membrane Microparticles From Plasma and Serum. Blood 60, no. 4 (1982): 834–840.
    pubmed: 7115953
  47. Palviainen M, Saraswat M, Varga Z. Extracellular Vesicles From Human Plasma and Serum Are Carriers of Extravesicular Cargo—Implications for Biomarker Discovery. PLoS One 15, no. 8 (2020): e0236439.
    pmc: PMC7446890pubmed: 32813744
  48. Segura D, Monreal L, Pérez‐Pujol S. Assessment of Platelet Function in Horses: Ultrastructure, Flow Cytometry, and Perfusion Techniques. Journal of Veterinary Internal Medicine 20, no. 3 (2006): 581–588.
    pubmed: 16734093
  49. Reymond S, Gruaz L, Sanchez J C. Depletion of Abundant Plasma Proteins for Extracellular Vesicle Proteome Characterization: Benefits and Pitfalls. Analytical and Bioanalytical Chemistry 415, no. 16 (2023): 3177–3187.
    pmc: PMC10287573pubmed: 37069444
  50. Cooper T T, Dieters‐Castator D Z, Liu J. Targeted Proteomics of Plasma Extracellular Vesicles Uncovers MUC1 as Combinatorial Biomarker for the Early Detection of High‐Grade Serous Ovarian Cancer. Journal of Ovarian Research 17, no. 1 (2024): 149.
    pmc: PMC11253408pubmed: 39020428
  51. Zhao Z, Wijerathne H, Godwin A K, Soper S A. Erratum: Isolation and Analysis Methods of Extracellular Vesicles (EVs). Extracellular Vesicles and Circulating Nucleic Acids [Internet] (2021) [cited 2025 Dec 13].
    pmc: PMC11651119pubmed: 39697590
  52. Weng X, Cloutier G, Pibarot P, Durand L. Comparison and Simulation of Different Levels of Erythrocyte Aggregation With Pig, Horse, Sheep, Calf, and Normal Human Blood. Biorheology 33, no. 4–5 (1996): 365–377.
    pubmed: 8977661
  53. Furi I, Momen‐Heravi F, Szabo G. Extracellular Vesicle Isolation: Present and Future. Annals of Translational Medicine 5, no. 12 (2017): 263.
    pmc: PMC5497100pubmed: 28706931
  54. Esmaeili A, Baghaban Eslaminejad M, Hosseini S. Biomolecular Corona Potential in Extracellular Vesicle Engineering for Therapeutic Applications. Biomedicine & Pharmacotherapy 188 (2025): 118202.
    pubmed: 40418857
  55. Jankovičová J, Sečová P, Michalková K, Antalíková J. Tetraspanins, More Than Markers of Extracellular Vesicles in Reproduction. International Journal of Molecular Sciences 21, no. 20 (2020): 7568.
    pmc: PMC7589920pubmed: 33066349
  56. Willms E, Cabañas C, Mäger I, Wood M J A, Vader P. Extracellular Vesicle Heterogeneity: Subpopulations, Isolation Techniques, and Diverse Functions in Cancer Progression. Frontiers in Immunology 9 (2018): 738.
    pmc: PMC5936763pubmed: 29760691
  57. Fan Y, Pionneau C, Cocozza F. Differential Proteomics Argues Against a General Role for CD9, CD81 or CD63 in the Sorting of Proteins Into Extracellular Vesicles. Journal of Extracellular Vesicles 12, no. 8 (2023): 12352.
    pmc: PMC10390663pubmed: 37525398
  58. Okada‐Tsuchioka M, Kajitani N, Omori W, Kurashige T, Boku S, Takebayashi M. Tetraspanin Heterogeneity of Small Extracellular Vesicles in Human Biofluids and Brain Tissue. Biochemical and Biophysical Research Communications 627 (2022): 146–151.
    pubmed: 36037746
  59. Karimi N, Dalirfardouei R, Dias T, Lötvall J, Lässer C. Tetraspanins Distinguish Separate Extracellular Vesicle Subpopulations in Human Serum and Plasma – Contributions of Platelet Extracellular Vesicles in Plasma Samples. Journal of Extracellular Vesicles 11, no. 5 (2022): e12213.
    pmc: PMC9077141pubmed: 35524458
  60. Xiang H, Chen S, Zhou J, Guo J, Zhou Q. Characterization of Blood‐Derived Exosomal Proteins After Exercise. Journal of International Medical Research 48, no. 9 (2020): 0300060520957541.
    pmc: PMC7522842pubmed: 32972266
  61. Zubairova L D, Nabiullina R M, Nagaswami C. Circulating Microparticles Alter Formation, Structure and Properties of Fibrin Clots. Scientific Reports 5, no. 1 (2015): 17611.
    pmc: PMC4669434pubmed: 26635081
  62. Lea J, Sharma R, Yang F, Zhu H, Ward E S, Schroit A J. Detection of Phosphatidylserine‐Positive Exosomes as a Diagnostic Marker for Ovarian Malignancies: A Proof of Concept Study. Oncotarget 8, no. 9 (2017): 14395–14407.
    pmc: PMC5362413pubmed: 28122335
  63. Zhang Q, Jeppesen D K, Higginbotham J N. Supermeres Are Functional Extracellular Nanoparticles Replete With Disease Biomarkers and Therapeutic Targets. Nature Cell Biology 23, no. 12 (2021): 1240–1254.
    pmc: PMC8656144pubmed: 34887515
  64. Jeppesen D K, Zhang Q, Franklin J L, Coffey R J. Extracellular Vesicles and Nanoparticles: Emerging Complexities. Trends in Cell Biology 33, no. 8 (2023): 667–681.
    pmc: PMC10363204pubmed: 36737375
  65. Azkargorta M, Iloro I, Escobes I. Human Serum Extracellular Vesicle Proteomic Profile Depends on the Enrichment Method Employed. International Journal of Molecular Sciences 22, no. 20 (2021): 11144.
    pmc: PMC8540106pubmed: 34681804
  66. Vasilatis DM, Walker NJ, Borjesson DL. Amikacin Disaggregates Platelet Clumps in EDTA Blood Samples From Cats and Dogs When Added Postcollection. Veterinary Clinical Pathology 52, no. 2 (2023): 228–235.
    pubmed: 36849708
  67. Williams TL, Archer J. Effect of Prewarming EDTA Blood Samples to 37°C on Platelet Count Measured by Sysmex XT‐2000iV in Dogs, Cats, and Horses. Veterinary Clinical Pathology 45, no. 3 (2016): 444–449.
    pubmed: 27391303
  68. Hinchcliff KW, Kociba GJ, Mitten LA. Diagnosis of EDTA‐Dependent Pseudothrombocytopenia in a Horse. Journal of the American Veterinary Medical Association 203, no. 12 (1993): 1715–1716.
    pubmed: 8307824
  69. Kapoor KS, Harris K, Arian KA. High Throughput and Rapid Isolation of Extracellular Vesicles and Exosomes With Purity Using Size Exclusion Liquid Chromatography. Bioactive Materials 40 (2024): 683–695.
    pmc: PMC11407901pubmed: 39290685
  70. Ignjatovic V, Geyer PE, Palaniappan KK. Mass Spectrometry‐Based Plasma Proteomics: Considerations From Sample Collection to Achieving Translational Data. Journal of Proteome Research 18, no. 12 (2019): 4085–4097.
    pmc: PMC6898750pubmed: 31573204
  71. Brennan K, Martin K, FitzGerald SP. A Comparison of Methods for the Isolation and Separation of Extracellular Vesicles From Protein and Lipid Particles in Human Serum. Scientific Reports 10, no. 1 (2020): 1039.
    pmc: PMC6978318pubmed: 31974468
  72. Gaspar LS, Santana MM, Henriques C. Simple and Fast SEC‐Based Protocol to Isolate Human Plasma‐Derived Extracellular Vesicles for Transcriptional Research. Molecular Therapy 18 (2020): 723–737.
    pmc: PMC7452272pubmed: 32913880
  73. De Sousa KP, Rossi I, Abdullahi M, Ramirez MI, Stratton D, Inal JM. Isolation and Characterization of Extracellular Vesicles and Future Directions in Diagnosis and Therapy. Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology 15, no. 1 (2023): e1835.
    pmc: PMC10078256pubmed: 35898167
  74. Buschmann D, Kirchner B, Hermann S. Evaluation of Serum Extracellular Vesicle Isolation Methods for Profiling miRNAs by Next‐Generation Sequencing. Journal of Extracellular Vesicles 7, no. 1 (2018): 1481321.
    pmc: PMC5990937pubmed: 29887978

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,911 horse owners like you who receive our email updates on horse health and nutrition.