title: Positioning of bone marrow hematopoietic and stromal cells relative to blood flow in vivo: serially reconstituting hematopoietic stem cells reside in distinct nonperfused niches.
abstract: Hematopoietic stem cell (HSC) niches have been reported at the endosteum or adjacent to bone marrow (BM) vasculature. To investigate functional attributes of these niches, mice were perfused with Hoechst 33342 (Ho) in vivo before BM cell collection in presence of pump inhibitors and antibody stained. We report that the position of phenotypic HSCs, multipotent and myeloid progenitors relative to blood flow, follows a hierarchy reflecting differentiation stage, whereas mesenchymal stromal cells are perivascular. Furthermore, during granulocyte colony-stimulating factor-induced mobilization, HSCs migrated closer to blood flow, whereas stromal cells did not. Interestingly, phenotypic Lin(-)Sca1(+)KIT(+)CD41(-)CD48(-)CD150(+) HSCs segregated into 2 groups (Ho(neg) or Ho(med)), based on degree of blood/Ho perfusion of their niche. HSCs capable of serial transplantation and long-term bromodeoxyuridine label retention were enriched in Ho(neg) HSCs, whereas Ho(med) HSCs cycled more frequently and only reconstituted a single host. This suggests that the most potent HSC niches are enriched in locally secreted factors and low oxygen tension due to negligible blood flow. Importantly, blood perfusion of niches correlates better with HSC function than absolute distance from vasculature. This technique enables prospective isolation of serially reconstituting HSCs distinct from other less potent HSCs of the same phenotype, based on the in vivo niche in which they reside.
authors.affiliation: Biotherapy Program, Mater Medical Research Institute, South Brisbane, Australia.
authors.forename: Ingrid G
authors.initials: IG
authors.lastname: Winkler
authors.forename: Valérie
authors.initials: V
authors.lastname: Barbier
authors.forename: Robert
authors.initials: R
authors.lastname: Wadley
authors.forename: Andrew C W
authors.initials: AC
authors.lastname: Zannettino
authors.forename: Sharon
authors.initials: S
authors.lastname: Williams
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Lévesque
mesh_terms: D000818:Animals; D001562:Benzimidazoles; D001783:Blood Flow Velocity; D001853:Bone Marrow / Q000098:blood supply* / Q000187:drug effects; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology; D001973:Bromodeoxyuridine / Q000378:metabolism; D002454:Cell Differentiation; D015687:Cell Hypoxia; D005456:Fluorescent Dyes; D016179:Granulocyte Colony-Stimulating Factor / Q000008:administration & dosage; D019650:Hematopoietic Stem Cell Mobilization; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000502:physiology; D051379:Mice; D020297:Mice, Congenic; D008810:Mice, Inbred C57BL; D039902:Multipotent Stem Cells / Q000166:cytology / Q000502:physiology; D023461:Myeloid Progenitor Cells / Q000166:cytology / Q000502:physiology; D010641:Phenotype; D011994:Recombinant Proteins; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology
chemical_list: D001562:Benzimidazoles; D005456:Fluorescent Dyes; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D001973:Bromodeoxyuridine; C017807:bisbenzimide ethoxide trihydrochloride