eBook: Breakthroughs and Innovations in Immunology Research - 18

The derivation of large numbers of macrophages
and dendritic cells from bone marrow has become
a staple of researchers across several fields of immunology.
Our experiments help to characterize these
populations to provide a better understanding of
their phenotype, function, and abilities.
References
Figure 6. Positively isolated CX3CR1 cells were labelled with CFSE
and transferred into C57BL/6 mice via retro- orbital injection. On
day 3, the recipient mouse bone marrow was isolated and stained
for CD11b, F4/80, and CX3CR1.
and dendritic cells. Realizing that these cells share
overlapping markers, it benefits researchers to look
at more phenotypic characteristics, such as comprehensive
cytokine and chemokine expression profile,
upon pattern recognition receptors stimulation,
and other critical stimuli. This is particularly true of
bone marrow CX3CR1+
precursors, as less is understood
about them compared to their peripheral
counterparts. Following isolation of these cells with
MojoSort™, we differentiated them with GM-CSF
(macrophages) or GM-CSF/IL-4 (dendritic cells). We
analyzed surface expression of markers like CD80,
CD86, and MHC II, with each cell demonstrating
upregulation of costimulatory molecules following
stimulation with LPS or CpG. We also examined cytokine
production using LEGENDplex™. Each population
exhibited distinct cytokine profiles depending
on the type of TLR agonist used. Finally, we demonstrated
that CX3CR1+
cells are capable of migration
to the bone marrow after adoptive transfer in vivo.
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inflammatory bowel disease. Gastroenterology. 2007.
132(1):139-53.
4. Matheu MP et al. Generation of bone marrow derived
murine dendritic cells for use in 2-photon imaging. J Vis
Exp. 2008 Jul 9;(17). pii: 773.
5. Lehmann JS et al. Multiplex Cytokine Profiling of
Stimulated Mouse Splenocytes Using a Cytometric Beadbased
Immunoassay Platform. J Vis Exp. 2017 Nov 9;(129).
6. Ceredig R et al. Models of haematopoiesis: seeing the
wood for the trees. Nat Rev Immunol. 2009. 9:293-300.
7. Travlos GS. Normal Structure, Function, and Histology of
the Bone Marrow. Toxicol Pathol. 2006. 34(5):548.
8. Helft J et al. GM-CSF Mouse Bone Marrow Cultures
Comprise a Heterogeneous Population of CD11c(+)
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2015. 42(6):1197-211.
Additional Resource
Immunology
18

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