Flow cytometry analysis of immune cell subsets within the murine spleen, bone marrow, lymph nodes and synovial tissue in an Osteoarthritis model
Osteoarthritis (OA) is one of the most prevalent musculoskeletal diseases, affecting patients suffering from pain and physical limitations. Recent evidence indicates a potential inflammatory component of the disease, with both T-cells and monocytes/macrophages potentially associated with the pathoge...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article (Journal) Video |
| Language: | English |
| Published: |
4/24/2020
|
| In: |
JoVE. Video journal
Year: 2020, Issue: 158, Pages: 1-11 |
| ISSN: | 1940-087X |
| DOI: | 10.3791/61008 |
| Subjects: | |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.3791/61008 Verlag, lizenzpflichtig, Volltext: https://www.jove.com/video/61008/flow-cytometry-analysis-immune-cell-subsets-within-murine-spleen-bone 
|
| Author Notes: | Patrick Haubruck, Aimee C. Colbath, Yolanda Liu, Shihani Stoner, Cindy Shu, Christopher B. Little |
MARC
| LEADER | 00000cga a2200000 c 4500 | ||
|---|---|---|---|
| 001 | 1699769524 | ||
| 003 | DE-627 | ||
| 005 | 20230621150645.0 | ||
| 006 | m o | | | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 200604s2020 xx ||| | o ||eng c | ||
| 024 | 7 | |a 10.3791/61008 |2 doi | |
| 035 | |a (DE-627)1699769524 | ||
| 035 | |a (DE-599)KXP1699769524 | ||
| 035 | |a (OCoLC)1341328484 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rda | ||
| 041 | |a eng | ||
| 084 | |a 33 |2 sdnb | ||
| 100 | 1 | |a Haubruck, Patrick |d 1987- |e VerfasserIn |0 (DE-588)1052915507 |0 (DE-627)78929625X |0 (DE-576)408656670 |4 aut | |
| 245 | 1 | 0 | |a Flow cytometry analysis of immune cell subsets within the murine spleen, bone marrow, lymph nodes and synovial tissue in an Osteoarthritis model |c Patrick Haubruck, Aimee C. Colbath, Yolanda Liu, Shihani Stoner, Cindy Shu, Christopher B. Little |
| 264 | 1 | |c 4/24/2020 | |
| 300 | |a 1 Online-Ressource (1 Videodatei, 12:23 min) |b farbig | ||
| 300 | |a 11 | ||
| 336 | |a Text |b txt |2 rdacontent | ||
| 336 | |a zweidimensionales bewegtes Bild |b tdi |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Gesehen am 04.06.2020 | ||
| 500 | |a Enthält auch Textversion | ||
| 500 | |a Wissenschaftlicher Film. Deutschland. 2020 | ||
| 520 | |a Osteoarthritis (OA) is one of the most prevalent musculoskeletal diseases, affecting patients suffering from pain and physical limitations. Recent evidence indicates a potential inflammatory component of the disease, with both T-cells and monocytes/macrophages potentially associated with the pathogenesis of OA. Further studies postulated an important role for subsets of both inflammatory cell lineages, such as Th1, Th2, Th17, and T-regulatory lymphocytes, and M1, M2, and synovium-tissue-resident macrophages. However, the interaction between the local synovial and systemic inflammatory cellular response and the structural changes in the joint is unknown. To fully understand how T-cells and monocytes/macrophages contribute towards OA, it is important to be able to quantitively identify these cells and their subsets simultaneously in synovial tissue, secondary lymphatic organs and systemically (the spleen and bone marrow). Nowadays, the different inflammatory cell subsets can be identified by a combination of cell-surface markers making multi-color flow cytometry a powerful technique in investigating these cellular processes. In this protocol, we describe detailed steps regarding the harvest of synovial tissue and secondary lymphatic organs as well as generation of single cell suspensions. Furthermore, we present both an extracellular staining assay to identify monocytes/macrophages and their subsets as well as an extra- and intra-cellular staining assay to identify T-cells and their subsets within the murine spleen, bone marrow, lymph nodes and synovial tissue. Each step of this protocol was optimized and tested, resulting in a highly reproducible assay that can be utilized for other surgical and non-surgical OA mouse models. | ||
| 655 | 7 | |a Film |0 (DE-588)4017102-4 |0 (DE-627)104559683 |0 (DE-576)208918531 |2 gnd-content | |
| 700 | 1 | |a Colbath, Aimee C. |e VerfasserIn |4 aut | |
| 700 | 1 | |a Liu, Yolanda |e VerfasserIn |4 aut | |
| 700 | 1 | |a Stoner, Shihani |e VerfasserIn |4 aut | |
| 700 | 1 | |a Shu, Cindy |e VerfasserIn |4 aut | |
| 700 | 1 | |a Little, Christopher B. |e VerfasserIn |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t JoVE. Video journal |d [Erscheinungsort nicht ermittelbar] : [Verlag nicht ermittelbar], 2006 |g (2020), 158, Artikel-ID e61008, Seite 1-11 |h Online-Ressource |w (DE-627)521454778 |w (DE-600)2259946-0 |w (DE-576)338522980 |x 1940-087X |7 nngs |a Flow cytometry analysis of immune cell subsets within the murine spleen, bone marrow, lymph nodes and synovial tissue in an Osteoarthritis model |
| 773 | 1 | 8 | |g year:2020 |g number:158 |g elocationid:e61008 |g pages:1-11 |g extent:11 |a Flow cytometry analysis of immune cell subsets within the murine spleen, bone marrow, lymph nodes and synovial tissue in an Osteoarthritis model |
| 856 | 4 | 0 | |u https://doi.org/10.3791/61008 |x Verlag |x Resolving-System |z lizenzpflichtig |3 Volltext |
| 856 | 4 | 0 | |u https://www.jove.com/video/61008/flow-cytometry-analysis-immune-cell-subsets-within-murine-spleen-bone |x Verlag |z lizenzpflichtig |3 Volltext |
| 935 | |c vide | ||
| 937 | |a Wissenschaftlicher Film |b Deutschland |c 2020 | ||
| 951 | |a AR | ||
| 992 | |a 20200604 | ||
| 993 | |a Video | ||
| 994 | |a 2020 | ||
| 998 | |g 1052915507 |a Haubruck, Patrick |m 1052915507:Haubruck, Patrick |d 910000 |d 911460 |e 910000PH1052915507 |e 911460PH1052915507 |k 0/910000/ |k 1/910000/911460/ |p 1 |x j | ||
| 999 | |a KXP-PPN1699769524 |e 3682484043 | ||
| BIB | |a Y | ||
| SER | |a journal | ||
| JSO | |a {"name":{"displayForm":["Patrick Haubruck, Aimee C. Colbath, Yolanda Liu, Shihani Stoner, Cindy Shu, Christopher B. Little"]},"id":{"doi":["10.3791/61008"],"eki":["1699769524"]},"origin":[{"dateIssuedKey":"2020","dateIssuedDisp":"4/24/2020"}],"relHost":[{"disp":"Flow cytometry analysis of immune cell subsets within the murine spleen, bone marrow, lymph nodes and synovial tissue in an Osteoarthritis modelJoVE. Video journal","type":{"media":"Online-Ressource","bibl":"periodical"},"note":["Gesehen am 10.02.25","Inhaltliche Gliederung in: Behavior; Biochemistry; Bioengineering; Biology; Cancer research; Chemistry; Developmental biology; Engineering; Environment; Genetics; Immunology and infection; Medicine; Neuroscience"],"language":["eng"],"recId":"521454778","pubHistory":["1.2006 -"],"part":{"text":"(2020), 158, Artikel-ID e61008, Seite 1-11","extent":"11","year":"2020","pages":"1-11","issue":"158"},"titleAlt":[{"title":"JoVE journal"},{"title":"JoVE journal Biology"},{"title":"JoVE journal Medicine"},{"title":"JoVE journal Behavior"},{"title":"JoVE journal Biochemistry"},{"title":"JoVE journal Bioengineering"},{"title":"JoVE journal Cancer research"},{"title":"JoVE journal Chemistry"},{"title":"JoVE journal Developmental biology"},{"title":"JoVE journal Engineering"},{"title":"JoVE journal Environment"},{"title":"JoVE journal Genetics"},{"title":"JoVE journal Immunology and infection"},{"title":"JoVE journal Neuroscience"},{"title":"Journal of visualized experiments"}],"title":[{"partname":"Video journal","title_sort":"JoVE","title":"JoVE"}],"physDesc":[{"extent":"Online-Ressource"}],"origin":[{"publisherPlace":"[Erscheinungsort nicht ermittelbar]","publisher":"[Verlag nicht ermittelbar]","dateIssuedKey":"2006","dateIssuedDisp":"2006-"}],"id":{"issn":["1940-087X"],"zdb":["2259946-0"],"eki":["521454778"]}}],"physDesc":[{"extent":"11 S.","noteIll":"farbig"}],"person":[{"family":"Haubruck","given":"Patrick","roleDisplay":"VerfasserIn","display":"Haubruck, Patrick","role":"aut"},{"role":"aut","roleDisplay":"VerfasserIn","display":"Colbath, Aimee C.","given":"Aimee C.","family":"Colbath"},{"given":"Yolanda","family":"Liu","role":"aut","display":"Liu, Yolanda","roleDisplay":"VerfasserIn"},{"family":"Stoner","given":"Shihani","roleDisplay":"VerfasserIn","display":"Stoner, Shihani","role":"aut"},{"roleDisplay":"VerfasserIn","display":"Shu, Cindy","role":"aut","family":"Shu","given":"Cindy"},{"given":"Christopher B.","family":"Little","role":"aut","roleDisplay":"VerfasserIn","display":"Little, Christopher B."}],"title":[{"title_sort":"Flow cytometry analysis of immune cell subsets within the murine spleen, bone marrow, lymph nodes and synovial tissue in an Osteoarthritis model","title":"Flow cytometry analysis of immune cell subsets within the murine spleen, bone marrow, lymph nodes and synovial tissue in an Osteoarthritis model"}],"language":["eng"],"recId":"1699769524","type":{"media":"Online-Ressource","bibl":"article-journal"},"note":["Gesehen am 04.06.2020","Enthält auch Textversion","Wissenschaftlicher Film. Deutschland. 2020"]} | ||
| SRT | |a HAUBRUCKPAFLOWCYTOME4242 | ||