MBTP 17: Immunophenotyping of dendritic cells from mouse spleen using flow cytometry

Miltenyi Biotec-tested panel 17 (MBTP 17)

This application protocol describes the flow cytometric analysis of dendritic cells (DCs) after spleen dissociation from healthy C57BL/6 mice. Mouse spleens are easily dissociated using the gentleMACS™ Dissociators to quickly obtain viable single cell suspensions ready for flow cytometric analysis.


Gating strategy showing the analysis of dendritic cell (DCs) populations from healthy mouse spleen. Splenocytes from C57BL/6 mice were stained using the previously described DC panel. Samples were initially gated on single cells using FSC-H/FSC-A gating and on live cells using Viobility-negative gating (data not shown). Conventional (CD11c+ MHC class II+) and plasmacytoid (Siglec-H+ CD317+) DCs were identified (A, D). Two major conventional DC (cDC) subsets, cDC1 and cDC2, can be found in spleen. The cDC1 subset is described as cross-presenting DC type and characteristically expresses XCR1 and lacks expression of CD172a. Non-cross presenting DCs (cDC2) express the opposite phenotype (XCR1 CD172a+) (B). Alternatively, cDC1 and cDC2 subsets can be differentiated using XCR1 and CD11b (C).


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