Detection and quantification methods of monocyte homing in coronary vasculature with an imaging cryomicrotome

Hakimzadeh, Nazanin; Horssen, Pepijn van; Lier, Monique G.J.T.B. van; Wijngaard, Jeroen P. H. M. van den; Belterman, Charly; Coronel, Ruben; Piek, Jan J.; Verberne, Hein J.; Spaan, Jos A. E.; Siebes, Maria

doi:10.1016/j.yjmcc.2014.08.019

Cited by 8 publications

(14 citation statements)

References 37 publications

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“…Total myocardial tissue volume was quantified using a binary representation of the myocardium obtained by applying a threshold according to the average background of the bright light reflection image stack. As described previously (11), fluorescently labeled autologous monocytes homed to the ischemic myocardium and appeared as clusters of bright intensity in the images. The tissue volume containing infiltrating autologous monocytes was quantified from binary images obtained by applying a threshold based on the average background intensity of the respective fluorescent label.…”

Section: Methodsmentioning

confidence: 72%

“…Progressive coronary artery narrowing was induced by implanting an ameroid constrictor (5.5 ϫ 1.5 mm; Research Instruments SW) on the first anterolateral branch of the left circumflex artery, as described previously (11). Maximal swelling of the ameroid material is reached in ϳ10 days.…”

Section: Methodsmentioning

confidence: 99%

“…Once monocytes were isolated into subpopulations (described below), cells were kept in culture overnight after which cells were labeled with live fluorescent cell trackers. CD14 ϩ CD62L ϩ monocytes were labeled with 25 M CellTracker Orange CMTMR (CMTMR), while CD14 ϩ CD62L Ϫ monocytes were labeled with 25 M CellTracker Red CMTPX (CMTPX; Invitrogen Molecular Probes), as described previously (11). Fluorescently labeled cell populations were combined and suspended in 10 ml phosphate-buffered saline (PBS; Lonza) with 0.4% heparin (Leo).…”

Section: Methodsmentioning

confidence: 99%

“…Monocytes were isolated from peripheral blood as described previously (11). After isolation of peripheral blood mononuclear cells (PBMCs) by Ficoll density gradient centrifugation, monocytes were probed with a mouse antihuman CD14-FITC monoclonal antibody (AbD Serotec; Clone TÜK4) and a biotinylated anti-human L-selectin (CD62L) antibody (R&D Systems), both at 1:20 dilution for 15 min at 4°C.…”

Section: Methodsmentioning

confidence: 99%

“…Sequential episcopic fluorescence image capture yielded multispectral 3D virtual reconstructions of the hearts with an in-plane resolution of 14 to 28 m. Isometric voxel size was achieved by average intensity projection of every two adjacent images in the stack. Processing of fluorescent vascular cast and cell images, including correction for spectral overlap of fluorescent cell tracker probes, was conducted as described previously (11,30).…”

Section: Methodsmentioning

confidence: 99%

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Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction

Hakimzadeh

Lier

Horssen

et al. 2016

American Journal of Physiology-Heart and Circulatory Physiology

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Hakimzadeh N, van Lier MG, van Horssen P, Daal M, Ly DH, Belterman C, Coronel R, Spaan JA, Siebes M. Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction. Am J Physiol Heart Circ Physiol 311: H239-H250, 2016. First published May 20, 2016 doi:10.1152/ajpheart.00187.2016.-Following myocardial infarction and atherosclerotic lesion development, monocytes contribute to myocardial protection and repair, while also partaking in myocardial ischemic injury. The balance of proinflammatory and reparative monocyte subsets is crucial in governing these therapeutic and pathological outcomes. Myocardial ischemic damage displays heterogeneity across the myocardium, whereby the subendocardium shows greatest vulnerability to ischemic damage. In this study we examined the transmural distribution of monocyte subsets in response to gradual coronary artery occlusion. CD14 ϩ monocytes were isolated from peripheral blood of New Zealand White rabbits and divided into two subgroups based on the expression of CD62L. We employed a rabbit model of progressive coronary artery obstruction to induce chronic myocardial ischemia and reinfused fluorescently labeled autologous monocytes. The distribution of fluorescently labeled autologous monocytes was examined with a high-resolution three-dimensional imaging cryomicrotome. The subepicardial layer contained the largest infiltration of both monocyte subgroups, with a significantly greater proportion of CD14 ϩ CD62L ϩ monocytes at the time when the ischemic area was at a maximum. By targeting CD13 ϩ angiogenic vessels, we confirmed the presence of angiogenesis in epicardial and midmyocardial regions. These myocardial regions demonstrated the highest level of infiltration of both monocyte subsets. Furthermore, CD14 ϩ CD62L ϩ monocytes showed significantly greater migration towards monocyte chemoattractant protein-1, greater adhesive capacity, and higher expression of C-C chemokine receptor type-2 relative to CD14 ϩ CD62L Ϫ monocytes. In conclusion, we note selective subepicardial distribution of monocyte subpopulations, with changes in proportion depending on the time after onset of coronary narrowing. Selective homing is supported by divergent migratory properties of each respective monocyte subgroup.

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Section: Methodsmentioning

confidence: 72%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction

Hakimzadeh

Lier

Horssen

et al. 2016

American Journal of Physiology-Heart and Circulatory Physiology

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Measurement and modeling of coronary blood flow

Sinclair

Lee

Cookson

et al. 2015

WIREs Mechanisms of Disease

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Ischemic heart disease that comprises both coronary artery disease and microvascular disease is the single greatest cause of death globally. In this context, enhancing our understanding of the interaction of coronary structure and function is not only fundamental for advancing basic physiology but also crucial for identifying new targets for treating these diseases. A central challenge for understanding coronary blood flow is that coronary structure and function exhibit different behaviors across a range of spatial and temporal scales. While experimental studies have sought to understand this feature by isolating specific mechanisms, in tandem, computational modeling is increasingly also providing a unique framework to integrate mechanistic behaviors across different scales. In addition, clinical methods for assessing coronary disease severity are continuously being informed and updated by findings in basic physiology. Coupling these technologies, computational modeling of the coronary circulation is emerging as a bridge between the experimental and clinical domains, providing a framework to integrate imaging and measurements from multiple sources with mathematical descriptions of governing physical laws. State-of-the-art computational modeling is being used to combine mechanistic models with data to provide new insight into coronary physiology, optimization of medical technologies, and new applications to guide clinical practice.

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Optimization of Vascular Casting for Three-Dimensional Fluorescence Cryo-Imaging of Collateral Vessels in the Ischemic Rat Hindlimb

et al. 2017

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Development of collateral vessels, arteriogenesis, may protect against tissue ischemia, however, quantitative data on this process remain scarce. We have developed a technique for replicating the entire arterial network of ischemic rat hindlimbs in three dimensions (3D) based on vascular casting and automated sequential cryo-imaging. Various dilutions of Batson's No. 17 with methyl methacrylate were evaluated in healthy rats, with further protocol optimization in ischemic rats. Penetration of the resin into the vascular network greatly depended on dilution; the total length of casted vessels below 75 µm was 13-fold higher at 50% dilution compared with the 10% dilution. Dilutions of 25-30%, with transient clamping of the healthy iliac artery, were optimal for imaging the arterial network in unilateral ischemia. This protocol completely filled the lumina of small arterioles and collateral vessels. These appeared as thin anastomoses in healthy legs and increasingly larger vessels during ligation (median diameter 1 week: 63 µm, 4 weeks: 127 µm). The presented combination of quality casts with high-resolution cryo-imaging enables automated, detailed 3D analysis of collateral adaptation, which furthermore can be combined with co-registered 3D distributions of fluorescent molecular imaging markers reflecting biological activity or perfusion.

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Detection and quantification methods of monocyte homing in coronary vasculature with an imaging cryomicrotome

Cited by 8 publications

References 37 publications

Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction

Selective subepicardial localization of monocyte subsets in response to progressive coronary artery constriction

Measurement and modeling of coronary blood flow

Optimization of Vascular Casting for Three-Dimensional Fluorescence Cryo-Imaging of Collateral Vessels in the Ischemic Rat Hindlimb

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