Guidelines for DC preparation and flow cytometry analysis of mouse nonlymphoid tissues

被引：8

作者：

Probst, Hans Christian ^{[1
,2
]}

Stoitzner, Patrizia ^{[3
]}

Amon, Lukas ^{[4
]}

Backer, Ronald A. ^{[2
,5
]}

Brand, Anna ^{[5
]}

Chen, Jianzhou ^{[6
]}

Clausen, Bjoern E. ^{[2
,5
]}

Dieckmann, Sophie ^{[3
]}

Dudziak, Diana ^{[7
,8
,9
]}

Heger, Lukas ^{[4
]}

Hodapp, Katrin ^{[2
]}

Hornsteiner, Florian ^{[3
]}

Hovav, Avi-Hai ^{[10
]}

Jacobi, Lukas ^{[4
]}

Ji, Xingqi ^{[11
,12
]}

Kamenjarin, Nadine ^{[2
]}

Lahl, Katharina ^{[13
,14
]}

Lahmar, Imran

Lakus, Jelena ^{[5
]}

Lehmann, Christian H. K. ^{[4
,7
]}

Ortner, Daniela ^{[3
]}

Picard, Marion ^{[6
]}

Roberti, Maria Paula ^{[6
,15
,16
]}

Rossnagel, Lukas ^{[4
]}

Saba, Yasmin ^{[10
]}

Schalla, Carmen ^{[17
,18
]}

Schlitzer, Andreas ^{[19
]}

Schraml, Barbara U. ^{[11
,12
]}

Schuetze, Kristian ^{[2
]}

Seichter, Anna ^{[4
]}

Sere, Kristin ^{[17
,18
]}

Seretis, Athanasios ^{[20
]}

Sopper, Sieghart ^{[21
,22
]}

Strandt, Helen ^{[3
]}

Sykora, Martina M. ^{[21
,22
]}

Theobald, Hannah ^{[19
]}

Tripp, Christoph H. ^{[3
]}

Zitvogel, Laurence ^{[6
]}

机构：

[1] Univ Med Ctr Mainz, Inst Immunol, Mainz, Germany

[2] Johannes Gutenberg Univ Mainz, Univ Med Ctr, Res Ctr Immunotherapy FZI, Mainz, Germany

[3] Med Univ Innsbruck, Dept Dermatol Venereol & Allergol, Innsbruck, Austria

[4] Univ Hosp Erlangen, Dept Dermatol, Lab Dendrit Cell Biol, Hartmannstr 14, D-91052 Erlangen, Germany

[5] Johannes Gutenberg Univ Mainz, Univ Med Ctr, Inst Mol Med, Paul Klein Ctr Immune Intervent, Mainz, Germany

[6] Univ Paris Saclay, GRCC, INSERM U1015, Villejuif, France

[7] Med Immunol Campus Erlangen MICE, D-91054 Erlangen, Germany

[8] DZI, Erlangen, Germany

[9] FAU, Erlangen, Germany

[10] Hebrew Univ Jerusalem, Inst Biomed & Oral Res, Fac Dent Med, Jerusalem, Israel

[11] Univ Hosp, Walter Brendel Ctr Expt Med, LMU Munich, D-82152 Planegg Martinsried, Germany

[12] Ludwig Maximilians Univ Munchen, Inst Cardiovasc Physiol & Pathophysiol, Fac Med, Biomed Ctr, D-82152 Martinsried, Germany

[13] Tech Univ Denmark DTU, Inst Hlth Technol, Sect Expt & Translat Immunol, DK-2800 Kongens Lyngby, Denmark

[14] Lund Univ, Immunol Sect, S-22184 Lund, Sweden

[15] Heidelberg Univ Hosp UKHD, Natl Ctr Tumor Dis NCT, Dept Med Oncol, Heidelberg, Germany

[16] German Canc Res Ctr, Clin Cooperat Unit Appl Tumor Immun, Heidelberg, Germany

[17] Rhein Westfal TH Aachen, Inst Biomed Engn, Dept Cell Biol, Med Sch, Aachen, Germany

[18] Rhein Westfal TH Aachen, Helmholtz Inst Biomed Engn, Aachen, Germany

[19] Univ Bonn, Quant Syst Biol Life & Med Sci LIMES Inst, Bonn, Germany

[20] Univ Innsbruck, Inst Biomed Aging Res, Innsbruck, Austria

[21] Med Univ Innsbruck, Internal Med 5, Hematol & Oncol, Innsbruck, Austria

[22] Tyrolean Canc Res Ctr, Innsbruck, Austria

来源：

EUROPEAN JOURNAL OF IMMUNOLOGY | 2022年

基金：

奥地利科学基金会; 欧洲研究理事会;

关键词：

Dendritic cells; Discrimination of dendritic cell subsets; Mouse nonlymphoid tissue; Multiplex flow cytometry analysis; Tissue digestion protocols; DENDRITIC CELL SUBSETS; HIGH-DIMENSIONAL ANALYSIS; DIVISION-OF-LABOR; FC-GAMMA-RIV; TRANSCRIPTIONAL CONTROL; LANGERHANS CELLS; STEADY-STATE; MACROPHAGES; MELANOMA; ANTIGEN;

D O I：

10.1002/eji.202249819

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

This article is part of the Dendritic Cell Guidelines article series, which provides a collection of state-of-the-art protocols for the preparation, phenotype analysis by flow cytometry, generation, fluorescence microscopy and functional characterization of mouse and human dendritic cells (DC) from lymphoid organs and various nonlymphoid tissues.DC are sentinels of the immune system present in almost every mammalian organ. Since they represent a rare cell population, DC need to be extracted from organs with protocols that are specifically developed for each tissue. This article provides detailed protocols for the preparation of single-cell suspensions from various mouse nonlymphoid tissues, including skin, intestine, lung, kidney, mammary glands, oral mucosa and transplantable tumors. Furthermore, our guidelines include comprehensive protocols for multiplex flow cytometry analysis of DC subsets and feature top tricks for their proper discrimination from other myeloid cells. With this collection, we provide guidelines for in-depth analysis of DC subsets that will advance our understanding of their respective roles in healthy and diseased tissues.While all protocols were written by experienced scientists who routinely use them in their work, this article was also peer-reviewed by leading experts and approved by all coauthors, making it an essential resource for basic and clinical DC immunologists.

引用

页数：78

共 142 条

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