Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids

Linda Bojmar; Han Sang Kim; Gabriel C. Tobias; Fanny A. Pelissier Vatter; Serena Lucotti; Kofi Ennu Gyan; Candia M. Kenific; Zurong Wan; Kyung A. Kim; Doo A. Kim; Jonathan Hernandez; Virginia Pascual; Todd E. Heaton; Michael P. La Quaglia; David Kelsen; Tanya M. Trippett; David R. Jones; William R. Jarnagin; Irina R. Matei; Haiying Zhang; Ayuko Hoshino; David Lyden

doi:10.1016/j.xpro.2020.100225

Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids

Linda Bojmar, Han Sang Kim, Gabriel C. Tobias, Fanny A. Pelissier Vatter, Serena Lucotti, Kofi Ennu Gyan, Candia M. Kenific, Zurong Wan, Kyung A. Kim, Doo A. Kim, Jonathan Hernandez, Virginia Pascual, Todd E. Heaton, Michael P. La Quaglia, David Kelsen, Tanya M. Trippett, David R. Jones, William R. Jarnagin, Irina R. Matei, Haiying ZhangAyuko Hoshino, David Lyden

Department of Internal Medicine

Research output: Contribution to journal › Article › peer-review

7 Citations (Scopus)

Abstract

We developed a modified protocol, based on differential ultracentrifugation (dUC), to isolate extracellular vesicles and particles (specifically exomeres) (EVPs) from various human and murine sources, including cell lines, surgically resected tumors and adjacent tissues, and bodily fluids, such as blood, lymphatic fluid, and bile. The diversity of these samples requires robust and highly reproducible protocols and refined isolation technology, such as asymmetric-flow field-flow fractionation (AF4). Our isolation protocol allows for preparation of EVPs for various downstream applications, including proteomic profiling. For complete details on the use and execution of this protocol, please refer to Hoshino et al. (2020).

Original language	English
Article number	100225
Journal	STAR Protocols
Volume	2
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2020.100225
Publication status	Published - 2021 Mar 19

Bibliographical note

Funding Information:
We thank the Electron Microscopy & Histology services of the Weill Cornell Medicine Microscopy & Image Analysis Core for TEM with NIH , United States Shared Instrumentation Grant ( S10RR027699 ) and Medical Illustration & Design, part of the Medical Research Support Services of Yonsei University College of Medicine , South Korea, for all artistic support related to this work. We are grateful for the great AF4 technical support from Wyatt Technology. The authors gratefully acknowledge support from: the National Cancer Institute , United States CA224175 (D.L.), CA210240 (D.L.), CA232093 (D.L.), CA163117 and CA207983 (D.L.), CA163120 (D.L.), CA169416 (D.L.), CA169538 (D.L.), CA218513 (D.L., H.Z.) and AI144301 (D.L., V.P.); the United States Department of Defense , United States W81XWH-13-1-0425 (D.L.), W81XWH-13-1-0427 , W81XWH-13-1-0249 (D.L.), and W81XWH-14-1-0199 (D.L.); National Institutes of Health , United States/WCM CTSC , United States (NIH/ NCATS ( UL1TR00457 ) (H.Z.); NIH/NCATS ( UL1TR002384 ) (D.L. and H.Z.); the Hartwell Foundation , United States (D.L.); the Thompson Family Foundation (D.L., D.K.); the STARR Consortium I9-A9-056 (D.L., H.Z.) and I8-A8-123 (D.L.); the Pediatric Oncology Experimental Therapeutics Investigator’s Consortium (D.L., T.M.T.); Alex’s Lemonade Stand Foundation , United Sates (D.L.); the Breast Cancer Research Foundation , United States (D.L.); the Feldstein Medical Foundation (D.L.); the Tortolani Foundation (D.L.); the Clinical & Translational Science Center (D.L., H.Z.); the Mary Kay Ash Charitable Foundation (D.L., I.M.); the Malcolm Hewitt Weiner Foundation (D.L.) ; the Manning Foundation (DL., A.H.) ; the Daniel P. and Nancy C. Paduano Family Foundation (D.L) ; the James Paduano Foundation (D.L.) ; the Sohn Foundation (D.L.) ; the AHEPA Vth District Cancer Research Foundation (D.L., L.B., S.L.) ; the Daedalus Fund Selma and Lawrence Ruben Science to Industry Bridge Award (D.L.); the Children’s Cancer and Blood Foundation , United States (D.L.); Susan G. Komen Postdoctoral Fellowship PDF15331556 , JST PRESTO, Japan 30021 , and JSPS KAKENHI, Japan JP19K23743 (A.H.); the National Research Foundation of Korea , South Korea (NRF) grant funded by the Korea government ( MSIT ) ( 2019R1C1C1006709 , 2018R1A5A2025079 , and 2020M3F7A1094093 ); a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare , South Korea ( KHIDIHI19C1015010020 ); "The Alchemist Project" through the Ministry of Trade, Industry and Energy (MOTIE, Korea) ( 20012443 ), and Severance Hospital Research fund for Clinical excellence (SHRC) ( C-2020-0032 and C-2020-0025 ) (H.S.K.); The Swedish Cancer Society Pancreatic Cancer Fellowship, the Lions International Postdoctoral fellowship and the Sweden-America stipend (L.B.); the São Paulo Research Foundation (FAPESP) São Paulo, Brazil ( 2017/07117-7 ) (G.C.T); the Swiss National Science Foundation (SFNS), Switzerland Postdoc Mobility grant ( P2SKP3_174785 and P400PB_186791 ) (F.A.V.); and the DoD PRCRP Horizon Award, United States ( W81XWH-19-PRCRP-HA ) (S.L).

Publisher Copyright:
© 2020 The Author(s)

All Science Journal Classification (ASJC) codes

Biochemistry, Genetics and Molecular Biology(all)
Neuroscience(all)
Immunology and Microbiology(all)
Medicine(all)

Access to Document

10.1016/j.xpro.2020.100225

Cite this

Bojmar, L., Kim, H. S., Tobias, G. C., Pelissier Vatter, F. A., Lucotti, S., Gyan, K. E., Kenific, C. M., Wan, Z., Kim, K. A., Kim, D. A., Hernandez, J., Pascual, V., Heaton, T. E., La Quaglia, M. P., Kelsen, D., Trippett, T. M., Jones, D. R., Jarnagin, W. R., Matei, I. R., ... Lyden, D. (2021). Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids. STAR Protocols, 2(1), [100225]. https://doi.org/10.1016/j.xpro.2020.100225

@article{4675a865e36c46b1a846526a051d6751,

title = "Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids",

abstract = "We developed a modified protocol, based on differential ultracentrifugation (dUC), to isolate extracellular vesicles and particles (specifically exomeres) (EVPs) from various human and murine sources, including cell lines, surgically resected tumors and adjacent tissues, and bodily fluids, such as blood, lymphatic fluid, and bile. The diversity of these samples requires robust and highly reproducible protocols and refined isolation technology, such as asymmetric-flow field-flow fractionation (AF4). Our isolation protocol allows for preparation of EVPs for various downstream applications, including proteomic profiling. For complete details on the use and execution of this protocol, please refer to Hoshino et al. (2020).",

author = "Linda Bojmar and Kim, {Han Sang} and Tobias, {Gabriel C.} and {Pelissier Vatter}, {Fanny A.} and Serena Lucotti and Gyan, {Kofi Ennu} and Kenific, {Candia M.} and Zurong Wan and Kim, {Kyung A.} and Kim, {Doo A.} and Jonathan Hernandez and Virginia Pascual and Heaton, {Todd E.} and {La Quaglia}, {Michael P.} and David Kelsen and Trippett, {Tanya M.} and Jones, {David R.} and Jarnagin, {William R.} and Matei, {Irina R.} and Haiying Zhang and Ayuko Hoshino and David Lyden",

note = "Funding Information: We thank the Electron Microscopy & Histology services of the Weill Cornell Medicine Microscopy & Image Analysis Core for TEM with NIH , United States Shared Instrumentation Grant ( S10RR027699 ) and Medical Illustration & Design, part of the Medical Research Support Services of Yonsei University College of Medicine , South Korea, for all artistic support related to this work. We are grateful for the great AF4 technical support from Wyatt Technology. The authors gratefully acknowledge support from: the National Cancer Institute , United States CA224175 (D.L.), CA210240 (D.L.), CA232093 (D.L.), CA163117 and CA207983 (D.L.), CA163120 (D.L.), CA169416 (D.L.), CA169538 (D.L.), CA218513 (D.L., H.Z.) and AI144301 (D.L., V.P.); the United States Department of Defense , United States W81XWH-13-1-0425 (D.L.), W81XWH-13-1-0427 , W81XWH-13-1-0249 (D.L.), and W81XWH-14-1-0199 (D.L.); National Institutes of Health , United States/WCM CTSC , United States (NIH/ NCATS ( UL1TR00457 ) (H.Z.); NIH/NCATS ( UL1TR002384 ) (D.L. and H.Z.); the Hartwell Foundation , United States (D.L.); the Thompson Family Foundation (D.L., D.K.); the STARR Consortium I9-A9-056 (D.L., H.Z.) and I8-A8-123 (D.L.); the Pediatric Oncology Experimental Therapeutics Investigator{\textquoteright}s Consortium (D.L., T.M.T.); Alex{\textquoteright}s Lemonade Stand Foundation , United Sates (D.L.); the Breast Cancer Research Foundation , United States (D.L.); the Feldstein Medical Foundation (D.L.); the Tortolani Foundation (D.L.); the Clinical & Translational Science Center (D.L., H.Z.); the Mary Kay Ash Charitable Foundation (D.L., I.M.); the Malcolm Hewitt Weiner Foundation (D.L.) ; the Manning Foundation (DL., A.H.) ; the Daniel P. and Nancy C. Paduano Family Foundation (D.L) ; the James Paduano Foundation (D.L.) ; the Sohn Foundation (D.L.) ; the AHEPA Vth District Cancer Research Foundation (D.L., L.B., S.L.) ; the Daedalus Fund Selma and Lawrence Ruben Science to Industry Bridge Award (D.L.); the Children{\textquoteright}s Cancer and Blood Foundation , United States (D.L.); Susan G. Komen Postdoctoral Fellowship PDF15331556 , JST PRESTO, Japan 30021 , and JSPS KAKENHI, Japan JP19K23743 (A.H.); the National Research Foundation of Korea , South Korea (NRF) grant funded by the Korea government ( MSIT ) ( 2019R1C1C1006709 , 2018R1A5A2025079 , and 2020M3F7A1094093 ); a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare , South Korea ( KHIDIHI19C1015010020 ); {"}The Alchemist Project{"} through the Ministry of Trade, Industry and Energy (MOTIE, Korea) ( 20012443 ), and Severance Hospital Research fund for Clinical excellence (SHRC) ( C-2020-0032 and C-2020-0025 ) (H.S.K.); The Swedish Cancer Society Pancreatic Cancer Fellowship, the Lions International Postdoctoral fellowship and the Sweden-America stipend (L.B.); the S{\~a}o Paulo Research Foundation (FAPESP) S{\~a}o Paulo, Brazil ( 2017/07117-7 ) (G.C.T); the Swiss National Science Foundation (SFNS), Switzerland Postdoc Mobility grant ( P2SKP3_174785 and P400PB_186791 ) (F.A.V.); and the DoD PRCRP Horizon Award, United States ( W81XWH-19-PRCRP-HA ) (S.L). Publisher Copyright: {\textcopyright} 2020 The Author(s)",

year = "2021",

month = mar,

day = "19",

doi = "10.1016/j.xpro.2020.100225",

language = "English",

volume = "2",

journal = "STAR Protocols",

issn = "2666-1667",

publisher = "Cell Press",

number = "1",

}

Bojmar, L, Kim, HS, Tobias, GC, Pelissier Vatter, FA, Lucotti, S, Gyan, KE, Kenific, CM, Wan, Z, Kim, KA, Kim, DA, Hernandez, J, Pascual, V, Heaton, TE, La Quaglia, MP, Kelsen, D, Trippett, TM, Jones, DR, Jarnagin, WR, Matei, IR, Zhang, H, Hoshino, A & Lyden, D 2021, 'Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids', STAR Protocols, vol. 2, no. 1, 100225. https://doi.org/10.1016/j.xpro.2020.100225

TY - JOUR

T1 - Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids

AU - Bojmar, Linda

AU - Kim, Han Sang

AU - Tobias, Gabriel C.

AU - Pelissier Vatter, Fanny A.

AU - Lucotti, Serena

AU - Gyan, Kofi Ennu

AU - Kenific, Candia M.

AU - Wan, Zurong

AU - Kim, Kyung A.

AU - Kim, Doo A.

AU - Hernandez, Jonathan

AU - Pascual, Virginia

AU - Heaton, Todd E.

AU - La Quaglia, Michael P.

AU - Kelsen, David

AU - Trippett, Tanya M.

AU - Jones, David R.

AU - Jarnagin, William R.

AU - Matei, Irina R.

AU - Zhang, Haiying

AU - Hoshino, Ayuko

AU - Lyden, David

N1 - Funding Information: We thank the Electron Microscopy & Histology services of the Weill Cornell Medicine Microscopy & Image Analysis Core for TEM with NIH , United States Shared Instrumentation Grant ( S10RR027699 ) and Medical Illustration & Design, part of the Medical Research Support Services of Yonsei University College of Medicine , South Korea, for all artistic support related to this work. We are grateful for the great AF4 technical support from Wyatt Technology. The authors gratefully acknowledge support from: the National Cancer Institute , United States CA224175 (D.L.), CA210240 (D.L.), CA232093 (D.L.), CA163117 and CA207983 (D.L.), CA163120 (D.L.), CA169416 (D.L.), CA169538 (D.L.), CA218513 (D.L., H.Z.) and AI144301 (D.L., V.P.); the United States Department of Defense , United States W81XWH-13-1-0425 (D.L.), W81XWH-13-1-0427 , W81XWH-13-1-0249 (D.L.), and W81XWH-14-1-0199 (D.L.); National Institutes of Health , United States/WCM CTSC , United States (NIH/ NCATS ( UL1TR00457 ) (H.Z.); NIH/NCATS ( UL1TR002384 ) (D.L. and H.Z.); the Hartwell Foundation , United States (D.L.); the Thompson Family Foundation (D.L., D.K.); the STARR Consortium I9-A9-056 (D.L., H.Z.) and I8-A8-123 (D.L.); the Pediatric Oncology Experimental Therapeutics Investigator’s Consortium (D.L., T.M.T.); Alex’s Lemonade Stand Foundation , United Sates (D.L.); the Breast Cancer Research Foundation , United States (D.L.); the Feldstein Medical Foundation (D.L.); the Tortolani Foundation (D.L.); the Clinical & Translational Science Center (D.L., H.Z.); the Mary Kay Ash Charitable Foundation (D.L., I.M.); the Malcolm Hewitt Weiner Foundation (D.L.) ; the Manning Foundation (DL., A.H.) ; the Daniel P. and Nancy C. Paduano Family Foundation (D.L) ; the James Paduano Foundation (D.L.) ; the Sohn Foundation (D.L.) ; the AHEPA Vth District Cancer Research Foundation (D.L., L.B., S.L.) ; the Daedalus Fund Selma and Lawrence Ruben Science to Industry Bridge Award (D.L.); the Children’s Cancer and Blood Foundation , United States (D.L.); Susan G. Komen Postdoctoral Fellowship PDF15331556 , JST PRESTO, Japan 30021 , and JSPS KAKENHI, Japan JP19K23743 (A.H.); the National Research Foundation of Korea , South Korea (NRF) grant funded by the Korea government ( MSIT ) ( 2019R1C1C1006709 , 2018R1A5A2025079 , and 2020M3F7A1094093 ); a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare , South Korea ( KHIDIHI19C1015010020 ); "The Alchemist Project" through the Ministry of Trade, Industry and Energy (MOTIE, Korea) ( 20012443 ), and Severance Hospital Research fund for Clinical excellence (SHRC) ( C-2020-0032 and C-2020-0025 ) (H.S.K.); The Swedish Cancer Society Pancreatic Cancer Fellowship, the Lions International Postdoctoral fellowship and the Sweden-America stipend (L.B.); the São Paulo Research Foundation (FAPESP) São Paulo, Brazil ( 2017/07117-7 ) (G.C.T); the Swiss National Science Foundation (SFNS), Switzerland Postdoc Mobility grant ( P2SKP3_174785 and P400PB_186791 ) (F.A.V.); and the DoD PRCRP Horizon Award, United States ( W81XWH-19-PRCRP-HA ) (S.L). Publisher Copyright: © 2020 The Author(s)

PY - 2021/3/19

Y1 - 2021/3/19

N2 - We developed a modified protocol, based on differential ultracentrifugation (dUC), to isolate extracellular vesicles and particles (specifically exomeres) (EVPs) from various human and murine sources, including cell lines, surgically resected tumors and adjacent tissues, and bodily fluids, such as blood, lymphatic fluid, and bile. The diversity of these samples requires robust and highly reproducible protocols and refined isolation technology, such as asymmetric-flow field-flow fractionation (AF4). Our isolation protocol allows for preparation of EVPs for various downstream applications, including proteomic profiling. For complete details on the use and execution of this protocol, please refer to Hoshino et al. (2020).

AB - We developed a modified protocol, based on differential ultracentrifugation (dUC), to isolate extracellular vesicles and particles (specifically exomeres) (EVPs) from various human and murine sources, including cell lines, surgically resected tumors and adjacent tissues, and bodily fluids, such as blood, lymphatic fluid, and bile. The diversity of these samples requires robust and highly reproducible protocols and refined isolation technology, such as asymmetric-flow field-flow fractionation (AF4). Our isolation protocol allows for preparation of EVPs for various downstream applications, including proteomic profiling. For complete details on the use and execution of this protocol, please refer to Hoshino et al. (2020).

UR - http://www.scopus.com/inward/record.url?scp=85103050917&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85103050917&partnerID=8YFLogxK

U2 - 10.1016/j.xpro.2020.100225

DO - 10.1016/j.xpro.2020.100225

M3 - Article

C2 - 33786456

AN - SCOPUS:85103050917

SN - 2666-1667

VL - 2

JO - STAR Protocols

JF - STAR Protocols

IS - 1

M1 - 100225

ER -

Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids

Abstract

Bibliographical note

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this