Deep imaging of bone marrow shows non-dividing stem cells are mainly perisinusoidal

Melih Acar, Kiranmai S. Kocherlakota, Malea M. Murphy, James G. Peyer, Hideyuki Oguro, Christopher N. Inra, Christabel Jaiyeola, Zhiyu Zhao, Katherine Luby-Phelps, Sean J. Morrison

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Abstract

Haematopoietic stem cells (HSCs) reside in a perivascular niche but the specific location of this niche remains controversial. HSCs are rare and few can be found in thin tissue sections or upon live imaging, making it difficult to comprehensively localize dividing and non-dividing HSCs. Here, using a green fluorescent protein (GFP) knock-in for the gene Ctnnal1 in mice (hereafter denoted as α-catulinGFP), we discover that α-catulinGFP is expressed by only 0.02% of bone marrow haematopoietic cells, including almost all HSCs. We find that approximately 30% of α-catulin-GFP+ c-kit+ cells give long-term multilineage reconstitution of irradiated mice, indicating that α-catulin-GFP+ c-kit+ cells are comparable in HSC purity to cells obtained using the best markers currently available. We optically cleared the bone marrow to perform deep confocal imaging, allowing us to image thousands of α-catulin-GFP+ c-kit+ cells and to digitally reconstruct large segments of bone marrow. The distribution of α-catulin-GFP+ c-kit+ cells indicated that HSCs were more common in central marrow than near bone surfaces, and in the diaphysis relative to the metaphysis. Nearly all HSCs contacted leptin receptor positive (Lepr+) and Cxcl12 high niche cells, and approximately 85% of HSCs were within 10 μm of a sinusoidal blood vessel. Most HSCs, both dividing (Ki-67+) and non-dividing (Ki-67-), were distant from arterioles, transition zone vessels, and bone surfaces. Dividing and non-dividing HSCs thus reside mainly in perisinusoidal niches with Lepr+ Cxcl12high cells throughout the bone marrow.

Original languageEnglish (US)
Pages (from-to)126-130
Number of pages5
JournalNature
Volume526
Issue number7571
DOIs
StatePublished - Oct 1 2015

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Hematopoietic Stem Cells
Stem Cells
Bone Marrow
Green Fluorescent Proteins
Bone Marrow Cells
Gene Knock-In Techniques
Leptin Receptors
Diaphyses
Arterioles
Blood Vessels
Bone and Bones

ASJC Scopus subject areas

  • General

Cite this

Acar, M., Kocherlakota, K. S., Murphy, M. M., Peyer, J. G., Oguro, H., Inra, C. N., ... Morrison, S. J. (2015). Deep imaging of bone marrow shows non-dividing stem cells are mainly perisinusoidal. Nature, 526(7571), 126-130. https://doi.org/10.1038/nature15250

Deep imaging of bone marrow shows non-dividing stem cells are mainly perisinusoidal. / Acar, Melih; Kocherlakota, Kiranmai S.; Murphy, Malea M.; Peyer, James G.; Oguro, Hideyuki; Inra, Christopher N.; Jaiyeola, Christabel; Zhao, Zhiyu; Luby-Phelps, Katherine; Morrison, Sean J.

In: Nature, Vol. 526, No. 7571, 01.10.2015, p. 126-130.

Research output: Contribution to journalArticle

Acar, M, Kocherlakota, KS, Murphy, MM, Peyer, JG, Oguro, H, Inra, CN, Jaiyeola, C, Zhao, Z, Luby-Phelps, K & Morrison, SJ 2015, 'Deep imaging of bone marrow shows non-dividing stem cells are mainly perisinusoidal', Nature, vol. 526, no. 7571, pp. 126-130. https://doi.org/10.1038/nature15250
Acar M, Kocherlakota KS, Murphy MM, Peyer JG, Oguro H, Inra CN et al. Deep imaging of bone marrow shows non-dividing stem cells are mainly perisinusoidal. Nature. 2015 Oct 1;526(7571):126-130. https://doi.org/10.1038/nature15250
Acar, Melih ; Kocherlakota, Kiranmai S. ; Murphy, Malea M. ; Peyer, James G. ; Oguro, Hideyuki ; Inra, Christopher N. ; Jaiyeola, Christabel ; Zhao, Zhiyu ; Luby-Phelps, Katherine ; Morrison, Sean J. / Deep imaging of bone marrow shows non-dividing stem cells are mainly perisinusoidal. In: Nature. 2015 ; Vol. 526, No. 7571. pp. 126-130.
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abstract = "Haematopoietic stem cells (HSCs) reside in a perivascular niche but the specific location of this niche remains controversial. HSCs are rare and few can be found in thin tissue sections or upon live imaging, making it difficult to comprehensively localize dividing and non-dividing HSCs. Here, using a green fluorescent protein (GFP) knock-in for the gene Ctnnal1 in mice (hereafter denoted as α-catulinGFP), we discover that α-catulinGFP is expressed by only 0.02{\%} of bone marrow haematopoietic cells, including almost all HSCs. We find that approximately 30{\%} of α-catulin-GFP+ c-kit+ cells give long-term multilineage reconstitution of irradiated mice, indicating that α-catulin-GFP+ c-kit+ cells are comparable in HSC purity to cells obtained using the best markers currently available. We optically cleared the bone marrow to perform deep confocal imaging, allowing us to image thousands of α-catulin-GFP+ c-kit+ cells and to digitally reconstruct large segments of bone marrow. The distribution of α-catulin-GFP+ c-kit+ cells indicated that HSCs were more common in central marrow than near bone surfaces, and in the diaphysis relative to the metaphysis. Nearly all HSCs contacted leptin receptor positive (Lepr+) and Cxcl12 high niche cells, and approximately 85{\%} of HSCs were within 10 μm of a sinusoidal blood vessel. Most HSCs, both dividing (Ki-67+) and non-dividing (Ki-67-), were distant from arterioles, transition zone vessels, and bone surfaces. Dividing and non-dividing HSCs thus reside mainly in perisinusoidal niches with Lepr+ Cxcl12high cells throughout the bone marrow.",
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