title: Quantitative imaging of haematopoietic stem and progenitor cell localization and hypoxic status in the bone marrow microenvironment.
abstract: The existence of a haematopoietic stem cell niche as a spatially confined regulatory entity relies on the notion that haematopoietic stem and progenitor cells (HSPCs) are strategically positioned in unique bone marrow microenvironments with defined anatomical and functional features. Here, we employ a powerful imaging cytometry platform to perform a comprehensive quantitative analysis of HSPC distribution in bone marrow cavities of femoral bones. We find that HSPCs preferentially localize in endosteal zones, where most closely interact with sinusoidal and non-sinusoidal bone marrow microvessels, which form a distinctive circulatory system. In situ tissue analysis reveals that HSPCs exhibit a hypoxic profile, defined by strong retention of pimonidazole and expression of HIF-1α, regardless of localization throughout the bone marrow, adjacency to vascular structures or cell-cycle status. These studies argue that the characteristic hypoxic state of HSPCs is not solely the result of a minimally oxygenated niche but may be partially regulated by cell-specific mechanisms.
authors.affiliation: Division of Transfusion Medicine, Department of Laboratory Medicine, Children's Hospital Boston, 300 Longwood Avenue, Boston, Massachusetts 02115, USA. Cesar.NombelaArrieta@childrens.harvard.edu [corrected]
authors.forename: César
authors.initials: C
authors.lastname: Nombela-Arrieta
authors.forename: Gregory
authors.initials: G
authors.lastname: Pivarnik
authors.forename: Beatrice
authors.initials: B
authors.lastname: Winkel
authors.forename: Kimberly J
authors.initials: KJ
authors.lastname: Canty
authors.forename: Brendan
authors.initials: B
authors.lastname: Harley
authors.forename: John E
authors.initials: JE
authors.lastname: Mahoney
authors.forename: Shin-Young
authors.initials: SY
authors.lastname: Park
authors.forename: Jiayun
authors.initials: J
authors.lastname: Lu
authors.forename: Alexei
authors.initials: A
authors.lastname: Protopopov
authors.forename: Leslie E
authors.initials: LE
authors.lastname: Silberstein
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000098:blood supply / Q000378:metabolism*; D002453:Cell Cycle; D015687:Cell Hypoxia; D060833:Cellular Microenvironment; D003114:Colony-Forming Units Assay; D005269:Femur / Q000166:cytology; D005434:Flow Cytometry / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D019394:Ki-67 Antigen / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D009593:Nitroimidazoles / Q000494:pharmacology; D013154:Spleen / Q000166:cytology / Q000378:metabolism; D019089:Stem Cell Factor / Q000378:metabolism
chemical_list: C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D019394:Ki-67 Antigen; C498355:Mki67 protein, mouse; D009593:Nitroimidazoles; D019089:Stem Cell Factor; C033815:pimonidazole