title: Bone-marrow-derived very small embryonic-like stem cells in patients with critical leg ischaemia: evidence of vasculogenic potential.
abstract: Very small embryonic-like stem cells (VSELs) are multipotent stem cells localised in adult bone marrow (BM) that may be mobilised into peripheral blood (PB) in response to tissue injury. We aimed to quantify VSELs in BM and PB of patients with critical limb ischaemia (CLI) and to test their angiogenic potential in vitro as well as their therapeutic capacity in mouse model of CLI. We isolated BM VSELs from patients with CLI and studied their potential to differentiate into vascular lineages. Flow and imaging cytometry showed that VSEL counts were lower in BM (p< 0.001) and higher (p< 0.001) in PB from CLI patients compared to healthy controls, suggesting that ischaemia may trigger VSELs mobilisation in this patient population. Sorted BM-VSELs cultured in angiogenic media acquired a mesenchymal phenotype (CD90+, Thy-1 gene positive expression). VSEL-derived cells had a pattern of secretion similar to that of endothelial progenitor cells, as they released low levels of VEGF-A and inflammatory cytokines. Noteworthy, VSELs triggered post-ischaemic revascularisation in immunodeficient mice (p< 0.05 vs PBS treatment), and acquired an endothelial phenotype either in vitro when cultured in the presence of VEGF-B (Cdh-5 gene positive expression), or in vivo in Matrigel implants (human CD31+ staining in neo-vessels from plug sections). In conclusion, VSELs are a potential new source of therapeutic cells that may give rise to cells of the endothelial lineage in humans.
authors.forename: Coralie L
authors.initials: CL
authors.lastname: Guerin
authors.forename: Xavier
authors.initials: X
authors.lastname: Loyer
authors.forename: José
authors.initials: J
authors.lastname: Vilar
authors.forename: Audrey
authors.initials: A
authors.lastname: Cras
authors.forename: Tristan
authors.initials: T
authors.lastname: Mirault
authors.forename: Pascale
authors.initials: P
authors.lastname: Gaussem
authors.forename: Jean-Sébastien
authors.initials: JS
authors.lastname: Silvestre
authors.affiliation: Prof. David Smadja, European Georges Pompidou Hospital, Hematology Department, 20 rue Leblanc, 75015 Paris, France, Tel.: +31 56093933, Fax: +31 56093393, E-mail: david.smadja@egp.aphp.fr.
authors.forename: David M
authors.initials: DM
authors.lastname: Smadja
mesh_terms: D000368:Aged; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation; D019070:Cell Lineage; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry; D005734:Gangrene / Q000628:therapy; D005786:Gene Expression Regulation; D006614:Hindlimb / Q000473:pathology; D006801:Humans; D007249:Inflammation; D007511:Ischemia / Q000628:therapy; D007869:Leg Injuries / Q000628:therapy*; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D009389:Neovascularization, Pathologic; D018919:Neovascularization, Physiologic; D010641:Phenotype; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D014456:Ulcer / Q000628:therapy*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D042561:Vascular Endothelial Growth Factor B / Q000378:metabolism
chemical_list: D019408:Platelet Endothelial Cell Adhesion Molecule-1; D042461:Vascular Endothelial Growth Factor A; D042561:Vascular Endothelial Growth Factor B
keywords: Endothelial cells; arterial thrombosis; bone marrow; stem cells; vasculogenesis