Imaging methods used to study mouse and human HSC niches: Current and emerging technologies

Tjin, Gavin; Flores-Figueroa, Eugenia; Duarte, Delfim; Straszkowski, Lenny; Scott, Marc; Khorshed, Reema; Purton, Louise E.; Celso, Cristina Lo

doi:10.1016/j.bone.2018.04.022

Cited by 21 publications

(19 citation statements)

References 104 publications

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“…In fact, the representativeness of BM TMA could strongly be affected by its tissue complexity and heterogeneity, even having the whole organ available. Particularly in humans, the selection of the core from BM trephines based on cellularity could exclude important regions i.e., those rich in adipocytes and/or bone trabecula (Tjin et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

High NESTIN Expression Marks the Endosteal Capillary Network in Human Bone Marrow

Panvini

Pacini

Montali

et al. 2020

Front. Cell Dev. Biol.

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Hematopoiesis is hosted, supported and regulated by a special bone marrow (BM) microenvironment known as “niche.” BM niches have been classified based on micro-anatomic distance from the bone surface into “endosteal” and “central” niches. Whilst different blood vessels have been found in both BM niches in mice, our knowledge of the human BM architecture is much more limited. Here, we have used a combination of markers including NESTIN, CD146, and αSMA labeling different blood vessels in benign human BM. Applying immunohistochemical/immunofluorescence techniques on BM trephines and performing image analysis on almost 300 microphotographs, we detected high NESTIN expression in BM endothelial cells (BMECs) of small arteries (A) and endosteal arterioles (EA), and also in very small vessels we named NESTIN+ capillary-like tubes (NCLTs), not surrounded by sub-endothelial perivascular cells that occasionally reported low levels of NESTIN expression. Statistically, NCLTs were detected within 40 μm from bone trabecula, frequently found in direct contact to the bone line and spatially correlated with hematopoietic stem/progenitor cells. Our results support the expression of NESTIN in human BMECs of EA and A in accordance with the updated classification of murine BM micro-vessels. NCLTs for their peculiar characteristics and micro-anatomical localization have been here proposed as transitional vessels possibly involved in regulating human hematopoiesis.

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Section: Introductionmentioning

confidence: 99%

High NESTIN Expression Marks the Endosteal Capillary Network in Human Bone Marrow

Panvini

Pacini

Montali

et al. 2020

Front. Cell Dev. Biol.

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“…Automatization of sample preparation and quantification is required to increase accuracy, comparability, and reproducibility of results (6). This is rapidly developing in clinical and experimental pathophysiology with image-analysis programs emerging for quantification and detection of specific cell types related to disease progression (42,43). Indeed, the high-quality image collection of over 192 murine H&E-stained bone sections that composes our dataset may become useful in training deep-learning algorithms for full automatization and finer detection of BM compartments.…”

Section: Discussionmentioning

confidence: 99%

MarrowQuant Across Aging and Aplasia: A Digital Pathology Workflow for Quantification of Bone Marrow Compartments in Histological Sections

et al. 2020

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in mid-sections of murine C57BL/6 bones in homeostatic conditions, including quantification of the highly predictable red-to-yellow transitions in the proximal section of the caudal tail and in the proximal-to-distal tibia. Additionally, we present a comparative skeletal map induced by lethal irradiation, with longitudinal quantification of the "red-to-yellow-to-red" transition over 2 months in C57BL/6 femurs and tibiae. We find that, following BM transplantation, BM adiposity inversely correlates with kinetics of hematopoietic recovery and that a proximal to distal gradient is conserved. Analysis of in vivo recovery through magnetic resonance imaging (MRI) reveals comparable kinetics. On human trephine biopsies MarrowQuant successfully recognizes the BM compartments, opening avenues for its application in experimental, or clinical contexts that require standardized human BM evaluation.

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“…Therefore, understanding homeostasis of the blood system requires knowledge of how the BM vascular network is organized and regulated. Intravital imaging of calvarial BM has helped define the spatiotemporal dynamics of BM vasculature and hematopoietic cells including HSPCs [17][18][19]26 . Three types of BM vasculature-arteries, arterioles, and sinusoids-contribute to blood homeostasis, although in different ways.…”

Section: Discussionmentioning

confidence: 99%

“…Others have extended this technology to analysis of the murine calvarium to assess migratory behavior of osteoclasts and spatiotemporal intercellular interactionsofliving osteoblasts and osteoclasts [13][14][15] . Calvarial bioimaging has resulted in important advances in hematopoiesis research, in part by defining positional relationships between HSCs and niche cells 16,17 . For example, combining fluorescent angiographic techniques with analysis in mouse reporter lines in which hematopoietic stem/progenitor cells (HSPCs) express a fluorescent protein has enabled us to trace hemodynamics and HSPC movement through vascular walls in the BM 18,19 .…”

Section: Physiological Regulation Of Blood Flow In Bone Marrow Is Impmentioning

confidence: 99%

Identification and local manipulation of bone marrow vasculature during intravital imaging

Morikawa

Tamaki

Fujita

et al. 2020

Sci Rep

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2 volumes PBS + 2%FCS, and centrifuged at 680 × g for 5 minutes. Cells were resuspended in PBS + 2%FCS and filtered through 40 μm nylon mesh. Cells were again centrifuged 680 × g for 5 minutes and treated with anti-CD16/32 antibody for Fc-receptor block (2 μL/mouse) for 5 minutes followed by addition of anti-CD45 magnetic beads (Miltenyi) at a 1/5 volume/volume ratio for 15 minutes. After removing the antibody with two PBS + 2%FCS washes, CD45-positive cells were isolated using Auto-MACS Pro (Miltenyi). Isolated cells were centrifuged once at 340 × g for 5 minutes. and injected into WT mice (3 × 10 6 cells/mouse). Statistical analysis. All values presented here are expressed as means ± SEM, unless noted otherwise. Differences between means were evaluated for significance using ANOVA followed by Fisher'sexact test for multiple comparisons. Differences with a p value < 0.05 were considered statistically significant.

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Imaging methods used to study mouse and human HSC niches: Current and emerging technologies

Cited by 21 publications

References 104 publications

High NESTIN Expression Marks the Endosteal Capillary Network in Human Bone Marrow

High NESTIN Expression Marks the Endosteal Capillary Network in Human Bone Marrow

MarrowQuant Across Aging and Aplasia: A Digital Pathology Workflow for Quantification of Bone Marrow Compartments in Histological Sections

Identification and local manipulation of bone marrow vasculature during intravital imaging

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