Mouse blood monocytes: Standardizing their identification and analysis using CD115

Breslin, Whitney L; Strohacker, Kelley; Carpenter, Katie C; Haviland, David L.; McFarlin, Brian K.

doi:10.1016/j.jim.2011.03.005

Cited by 38 publications

(23 citation statements)

References 27 publications

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“…Animal care and experimentation were performed accordingly with the Spanish Policy for Animal Protection RD53/2013, which meets the European Union Directive 2010/63/UE. The methodology allowed an accurate characterization of the cells in a small volume of whole peripheral blood, which is in accordance with previous studies (Breslin et al, ). Notwithstanding, earlier stages than P30 were not manageable to avoid animal suffering and stress due to their small size.…”

Section: Methodssupporting

confidence: 88%

Comparative study of hematopoietic stem and progenitor cells between sexes in mice under physiological conditions along time

Gasco

Rando

Zaragoza

et al. 2017

Cell Biology International

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Hematopoietic stem and progenitor cells (HSPCs) are attractive targets in regenerative medicine, although the differences in their homeostatic maintenance between sexes along time are still under debate. We accurately monitored hematopoietic stem cells (HSCs), common lymphoid progenitors (CLPs), and common myeloid progenitors (CMPs) frequencies by flow cytometry, by performing serial peripheral blood extractions from male and female B6SJL wild-type mice and found no significant differences. Only modest differences were found in the gene expression profile of Slamf1 and Gata2. Our findings suggest that both sexes could be used indistinctly to perform descriptive studies in the murine hematopoietic system, especially for flow cytometry studies in peripheral blood. This would allow diminishing the number of animals needed for the experimental procedures. In addition, the use of serial extractions in the same animals drastically decreases the number of animals needed.

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Section: Methodssupporting

confidence: 88%

Comparative study of hematopoietic stem and progenitor cells between sexes in mice under physiological conditions along time

Gasco

Rando

Zaragoza

et al. 2017

Cell Biology International

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“…The identity of these cells as circulatory monocytes was further confirmed by staining for CD115 (Panel a of Supplementary Fig. IV), a well-known marker of murine blood monocytes 30 . We also found significantly enhanced infiltrations of highly inflammatory Gr1-Mac2 (~2-fold, P<0.006, Fig.…”

Section: Resultsmentioning

confidence: 71%

Ribosomal Protein L13a Deficiency in Macrophages Promotes Atherosclerosis by Limiting Translation Control-Dependent Retardation of Inflammation

Basu

Poddar

Robinet³

et al. 2014

ATVB

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Objective Unresolved inflammatory response of macrophages plays a pivotal role in the pathogenesis of atherosclerosis. Previously we showed that ribosomal protein L13a-dependent translational silencing suppresses the synthesis of a cohort of inflammatory proteins in monocytes and macrophages. We also found that genetic abrogation of L13a expression in macrophages significantly compromised the resolution of inflammation in a mouse model of LPS-induced endotoxemia. However, its function in the pathogenesis of atherosclerosis is not known. Here, we examine whether L13a in macrophage has a protective role against high fat diet-induced atherosclerosis. Approach and Results We bred the macrophage-specific L13a knockout mice L13a Flox+/+ Cre+/+ onto apoE−/− background and generated the experimental double knockout (KO) mice L13a Flox+/+ Cre+/+ apoE−/−. L13a Flox+/+ Cre−/− mice on apoE−/− background were used as controls. Control and KO mice were subjected to high-fat diet for 10 weeks. Evaluation of aortic sinus sections and entire aorta by en face showed significantly higher atherosclerosis in the KO mice. Severity of atherosclerosis in KO mice was accompanied by thinning of the smooth muscle cell (SMC) layer in the media, larger macrophage area in the intimal plaque region and higher plasma levels of inflammatory cytokines. In addition, macrophages isolated from KO mice had higher polyribosomal abundance of several target mRNAs, thus showing defect in translation control. Conclusion Our data demonstrate that loss of L13a in macrophages increases susceptibility to atherosclerosis in apoE−/− mice, revealing an important role of L13a-dependent translational control as an endogenous protection mechanism against atherosclerosis.

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“…Considering that the average body weight of the population of 180 C57BL/6 mice analyzed in our study was 25 g, the maximum daily blood sample volume should not exceed 20 µl (National Society for Laboratory Animal Science, 2009). On the other hand, as shown previously, using blood amounts below 20 µl could hinder the proper differentiation between monocyte subsets [16]. We therefore compared the concentration of monocyte and granulocyte subsets between assay-standardized (50 µl) and volume-reduced (20 µl) blood samples (Figure 1 B).…”

Section: Resultsmentioning

confidence: 95%

Sham Surgery and Inter-Individual Heterogeneity Are Major Determinants of Monocyte Subset Kinetics in a Mouse Model of Myocardial Infarction

et al. 2014

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AimsMouse models of myocardial infarction (MI) are commonly used to explore the pathophysiological role of the monocytic response in myocardial injury and to develop translational strategies. However, no study thus far has examined the potential impact of inter-individual variability and sham surgical procedures on monocyte subset kinetics after experimental MI in mice. Our goal was to investigate determinants of systemic myeloid cell subset shifts in C57BL/6 mice following MI by developing a protocol for sequential extensive flow cytometry (FCM).Methods and ResultsFollowing cross-sectional multiplex FCM analysis we provide for the first time a detailed description of absolute quantities, relative subset composition, and biological variability of circulating classical, intermediate, and non-classical monocyte subsets in C57BL/6 mice. By using intra-individual longitudinal measurements after MI induction, a time course of classical and non-classical monocytosis was recorded. This approach disclosed a significant reduction of monocyte subset dispersion across all investigated time points following MI. We found that in the current invasive model of chronic MI the global pattern of systemic monocyte kinetics is mainly determined by a nonspecific inflammatory response to sham surgery and not by the extent of myocardial injury.ConclusionsApplication of sequential multiplexed FCM may help to reduce the impact of biological variability in C57BL/6 mice. Furthermore, the confounding influence of sham surgical procedures should always be considered when measuring monocyte subset kinetics in a murine model of MI.

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Mouse blood monocytes: Standardizing their identification and analysis using CD115

Cited by 38 publications

References 27 publications

Comparative study of hematopoietic stem and progenitor cells between sexes in mice under physiological conditions along time

Comparative study of hematopoietic stem and progenitor cells between sexes in mice under physiological conditions along time

Ribosomal Protein L13a Deficiency in Macrophages Promotes Atherosclerosis by Limiting Translation Control-Dependent Retardation of Inflammation

Sham Surgery and Inter-Individual Heterogeneity Are Major Determinants of Monocyte Subset Kinetics in a Mouse Model of Myocardial Infarction

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