Identification of the adult human hemangioblast.
Recent studies show that human CD133(+) (previously known as AC133(+)) cells from mobilized peripheral blood consist of stem cells with either hematopoietic or endothelial potential. To test whether this population also contains individual precursors with both capacities, the defining characteristics of the elusive adult hemangioblast, we developed a culture system that allows single-cell analyses of differentiation. In the presence of vascular endothelial growth factor (VEGF), stem cell growth factor (SCGF), and FLT-3 ligand, CD133(+)-enriched cells were first expanded and the amplified cells were transduced with a vector encoding an enhanced green fluorescent protein (EGFP) marker gene. Single EGFP(+) cells were then cocultured with corresponding non-transduced cells from the same experiment, yielding 50-100 marked cells in 8% of the wells after 2 weeks. The resultant cells were divided and differentiated with either granulocyte colony-stimulating factor (G-CSF) or with SCGF and VEGF. These culture conditions resulted in the formation of neutrophil or endothelial cells, respectively, as identified morphologically and by phenotypic staining. Dual differentiation of EGFP(+) cells could be observed in one-quarter of clones from single-seeded cells, suggesting that 2% of EGFP(+) cells were in fact human hemangioblasts. These cells could be expanded for at least 28 days without losing this dual capacity. Hence, this culture system may be of clinical relevance in the development of cellular therapies for disorders involving hematopoiesis and the vascular system. In addition, our results provide important information related to the development of the vasculature and the potential role of hemangioblasts in vasculogenesis in adult humans.
Loges
Sonja
S
Department of Medicine, University Hospital Eppendorf, Hamburg, Germany.
Fehse
Boris
B
Brockmann
Marc A
MA
Lamszus
Katrin
K
Butzal
Martin
M
Guckenbiehl
Markus
M
Schuch
Gunter
G
Ergün
Süleyman
S
Fischer
Uta
U
Zander
Axel R
AR
Hossfeld
Dieter K
DK
Fiedler
Walter
W
Gehling
Ursula M
UM
D000071916:AC133 Antigen; D000328:Adult; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D017930:Genes, Reporter; D006023:Glycoproteins / Q000378:metabolism*; D016298:Hematopoietic Cell Growth Factors / Q000378:metabolism; D006801:Humans; D037181:Lectins, C-Type / Q000378:metabolism; D018919:Neovascularization, Physiologic; D010455:Peptides / Q000378:metabolism*; D010641:Phenotype; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014161:Transduction, Genetic; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D051941:fms-Like Tyrosine Kinase 3 / Q000378:metabolism
D000071916:AC133 Antigen; D015703:Antigens, CD; C110532:CLEC11A protein, human; D016207:Cytokines; D006023:Glycoproteins; D016298:Hematopoietic Cell Growth Factors; D037181:Lectins, C-Type; C000606023:PROM1 protein, human; D010455:Peptides; D042461:Vascular Endothelial Growth Factor A; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3