Mobilization of hematopoietic stem cells by depleting bone marrow macrophages.
An important factor contributing to hematopoietic stem cell (HSC) mobilization is the ability of mobilizing cytokines and chemotherapy to disturb the cellular components of HSC niches, particularly osteoblasts and their progenitors, and to inhibit the production of HSC supportive cytokines and chemokines. Although the mechanisms by which niche cells are inhibited by mobilizing treatments is still incompletely understood, it has recently emerged that bone marrow macrophages play a critical role in maintaining osteoblasts, bone formation, and the expression of CXCL12, KIT ligand, and angiopoietin-1 necessary to HSC maintenance. In this chapter, we describe how to mobilize HSC into the blood in mice by depleting macrophages with clodronate-loaded liposomes and compare this mode of mobilization to mobilization induced by granulocyte colony-stimulating factor and cyclophosphamide. Detailed methods to analyze mobilization of phenotypic and functional reconstituting HSC are described with examples.
Mater Medical Research Institute, Aubigny Place, Raymond Terrace, South Brisbane, QLD, Australia.
Valérie
V
Barbier
Ingrid G
IG
Winkler
Jean-Pierre
JP
Lévesque
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects; D004002:Clodronic Acid / Q000494:pharmacology; D003114:Colony-Forming Units Assay; D003520:Cyclophosphamide / Q000494:pharmacology; D005260:Female; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D019650:Hematopoietic Stem Cell Mobilization / Q000379:methods*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects; D016130:Immunophenotyping; D047589:Leukocyte Reduction Procedures / Q000379:methods*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D008297:Male; D051379:Mice
D004002:Clodronic Acid; D016179:Granulocyte Colony-Stimulating Factor; D003520:Cyclophosphamide