Comparison of Methods for Isolation and Characterization of Total and Astrocyte‐Enriched Extracellular Vesicles From Human Serum and Plasma
ABSTRACT Extracellular vesicles (EV) which play critical roles in intercellular communication, have garnered interest as biomarkers with researchers studying brain‐related disease processes due to their ability to be isolated from various biofluids. Astrocytes, a type of glial cell, play a critical...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2025-02-01
|
| Series: | Journal of Extracellular Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/jex2.70035 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850079916275531776 |
|---|---|
| author | Leandra K. Figueroa‐Hall Kaiping Burrows Ahlam M. Alarbi Bethany N. Hannafon Cole Hladik Chibing Tan Rajagopal Ramesh Jennifer L. Stewart Victoria B. Risbrough Martin P. Paulus T. Kent Teague |
| author_facet | Leandra K. Figueroa‐Hall Kaiping Burrows Ahlam M. Alarbi Bethany N. Hannafon Cole Hladik Chibing Tan Rajagopal Ramesh Jennifer L. Stewart Victoria B. Risbrough Martin P. Paulus T. Kent Teague |
| author_sort | Leandra K. Figueroa‐Hall |
| collection | DOAJ |
| description | ABSTRACT Extracellular vesicles (EV) which play critical roles in intercellular communication, have garnered interest as biomarkers with researchers studying brain‐related disease processes due to their ability to be isolated from various biofluids. Astrocytes, a type of glial cell, play a critical role in neuronal regulation and function. As such, EV enriched from astrocytes can be used to interrogate cargo and identify mechanisms by which astrocytes communicate with other cells of the central nervous system or shed light on pathophysiological conditions. This manuscript compared five EV isolation methods (differential ultracentrifugation [dUC], precipitation, precipitation + purification, silicon carbon resin and size exclusion chromatography [SEC]) using small volumes of human plasma and serum with a focus on immunocapture of astrocyte‐enriched EV (AEEV), with the excitatory amino acid transporter 1, or GLAST. Methods were evaluated on yield, purity, recovery and downstream application to include immunoassays for tetraspanin, immune and astrocyte markers. Results revealed that whilst precipitation‐based methods such as ExoQuick yielded higher EV concentrations, size exclusion (SmartSEC, qEV) provided greater purity, emphasizing a trade‐off between yield and purity. This study provides a comprehensive resource for researchers in selecting EV isolation methods tailored to small biobanked clinical samples, with the goal of advancing biomarker discovery in Neuroscience. |
| format | Article |
| id | doaj-art-d1b90894a8964d9ab1dcb70f581fb5d4 |
| institution | DOAJ |
| issn | 2768-2811 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Extracellular Biology |
| spelling | doaj-art-d1b90894a8964d9ab1dcb70f581fb5d42025-08-20T02:45:04ZengWileyJournal of Extracellular Biology2768-28112025-02-0142n/an/a10.1002/jex2.70035Comparison of Methods for Isolation and Characterization of Total and Astrocyte‐Enriched Extracellular Vesicles From Human Serum and PlasmaLeandra K. Figueroa‐Hall0Kaiping Burrows1Ahlam M. Alarbi2Bethany N. Hannafon3Cole Hladik4Chibing Tan5Rajagopal Ramesh6Jennifer L. Stewart7Victoria B. Risbrough8Martin P. Paulus9T. Kent Teague10Laureate Institute for Brain Research Tulsa Oklahoma USALaureate Institute for Brain Research Tulsa Oklahoma USAIntegrative Immunology Center University of Oklahoma (OU) Tulsa Oklahoma USADepartment of Obstetrics and Gynecology OU Health Sciences Center (OUHSC) Oklahoma City Oklahoma USADepartment of Obstetrics and Gynecology OU Health Sciences Center (OUHSC) Oklahoma City Oklahoma USAIntegrative Immunology Center University of Oklahoma (OU) Tulsa Oklahoma USAOU Health Stephenson Cancer Center Oklahoma City Oklahoma USALaureate Institute for Brain Research Tulsa Oklahoma USAVA Center of Excellence for Stress and Mental Health La Jolla California USALaureate Institute for Brain Research Tulsa Oklahoma USAIntegrative Immunology Center University of Oklahoma (OU) Tulsa Oklahoma USAABSTRACT Extracellular vesicles (EV) which play critical roles in intercellular communication, have garnered interest as biomarkers with researchers studying brain‐related disease processes due to their ability to be isolated from various biofluids. Astrocytes, a type of glial cell, play a critical role in neuronal regulation and function. As such, EV enriched from astrocytes can be used to interrogate cargo and identify mechanisms by which astrocytes communicate with other cells of the central nervous system or shed light on pathophysiological conditions. This manuscript compared five EV isolation methods (differential ultracentrifugation [dUC], precipitation, precipitation + purification, silicon carbon resin and size exclusion chromatography [SEC]) using small volumes of human plasma and serum with a focus on immunocapture of astrocyte‐enriched EV (AEEV), with the excitatory amino acid transporter 1, or GLAST. Methods were evaluated on yield, purity, recovery and downstream application to include immunoassays for tetraspanin, immune and astrocyte markers. Results revealed that whilst precipitation‐based methods such as ExoQuick yielded higher EV concentrations, size exclusion (SmartSEC, qEV) provided greater purity, emphasizing a trade‐off between yield and purity. This study provides a comprehensive resource for researchers in selecting EV isolation methods tailored to small biobanked clinical samples, with the goal of advancing biomarker discovery in Neuroscience.https://doi.org/10.1002/jex2.70035astrocyte‐enriched extracellular vesiclesdifferential centrifugationextracellular vesiclesprecipitationprecipitation + purificationresin‐separating matrix |
| spellingShingle | Leandra K. Figueroa‐Hall Kaiping Burrows Ahlam M. Alarbi Bethany N. Hannafon Cole Hladik Chibing Tan Rajagopal Ramesh Jennifer L. Stewart Victoria B. Risbrough Martin P. Paulus T. Kent Teague Comparison of Methods for Isolation and Characterization of Total and Astrocyte‐Enriched Extracellular Vesicles From Human Serum and Plasma Journal of Extracellular Biology astrocyte‐enriched extracellular vesicles differential centrifugation extracellular vesicles precipitation precipitation + purification resin‐separating matrix |
| title | Comparison of Methods for Isolation and Characterization of Total and Astrocyte‐Enriched Extracellular Vesicles From Human Serum and Plasma |
| title_full | Comparison of Methods for Isolation and Characterization of Total and Astrocyte‐Enriched Extracellular Vesicles From Human Serum and Plasma |
| title_fullStr | Comparison of Methods for Isolation and Characterization of Total and Astrocyte‐Enriched Extracellular Vesicles From Human Serum and Plasma |
| title_full_unstemmed | Comparison of Methods for Isolation and Characterization of Total and Astrocyte‐Enriched Extracellular Vesicles From Human Serum and Plasma |
| title_short | Comparison of Methods for Isolation and Characterization of Total and Astrocyte‐Enriched Extracellular Vesicles From Human Serum and Plasma |
| title_sort | comparison of methods for isolation and characterization of total and astrocyte enriched extracellular vesicles from human serum and plasma |
| topic | astrocyte‐enriched extracellular vesicles differential centrifugation extracellular vesicles precipitation precipitation + purification resin‐separating matrix |
| url | https://doi.org/10.1002/jex2.70035 |
| work_keys_str_mv | AT leandrakfigueroahall comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma AT kaipingburrows comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma AT ahlammalarbi comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma AT bethanynhannafon comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma AT colehladik comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma AT chibingtan comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma AT rajagopalramesh comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma AT jenniferlstewart comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma AT victoriabrisbrough comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma AT martinppaulus comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma AT tkentteague comparisonofmethodsforisolationandcharacterizationoftotalandastrocyteenrichedextracellularvesiclesfromhumanserumandplasma |