Hematopoietic progenitor cells and interleukin-stimulated endothelium: expansion and differentiation of myeloid precursors.
BACKGROUND
Cytokine-stimulated endothelial cells (EC) propagate hematopoietic progenitor cell (HPC) expansion. However, the effects on the functional capacities of cultured progenitors have not been evaluated. HPC were assessed by flow cytometry, colony and cobblestone assays and long-term cultures (LTC) after culturing in the supernatant of EC stimulated by IL-1beta, IL-3 or IL-6.


RESULTS
EC incubation with IL-6 did not improve cell expansion in comparison to non-stimulated EC supernatant, while the HPCs' phenotype and functional capacities were retained. In contrast, IL-1beta and IL-3 stimulation resulted in a 10- and 100-fold increase in cell numbers with more than 90% of these cells being CD33(+). Plating efficiencies and LTC initiating cells were greatest in IL-6 supernatants, whereas the highest numbers of burst-forming units were observed using IL-3. IL-1beta supernatants diminished the number of 5-week cobblestone-areas, whereas the number of 2-week cobblestone areas remained equal to freshly isolated HPC. Fewer 2-week cobblestones and greater amounts of 5-week cobblestones were observed with IL-6 and IL-3. Expanded progenitors from all interleukin conditions were further matured into functional granulocytes.


CONCLUSIONS
IL-1beta and IL-3 stimulated endothelium induces proliferation and differentiation of myeloid precursors, while IL-6 treatment induced a benefit of HPC survival.
Moldenhauer
Anja
A
Institute for Transfusion Medicine, Charité - Universitätsmedizin Berlin, Germany. amolden@charite.de
Genter
Gesche
G
Lun
Andreas
A
Bal
Gürkan
G
Kiesewetter
Holger
H
Salama
Abdulgabar
A
D015703:Antigens, CD / Q000032:analysis; D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media; D042783:Endothelial Cells / Q000502:physiology*; D006098:Granulocytes / Q000166:cytology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D053583:Interleukin-1beta / Q000276:immunology; D007377:Interleukin-3 / Q000276:immunology; D015850:Interleukin-6 / Q000276:immunology; D007378:Interleukins / Q000276:immunology*; D023461:Myeloid Progenitor Cells / Q000166:cytology*; D063268:Sialic Acid Binding Ig-like Lectin 3; D014470:Umbilical Cord
D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; C571163:CD33 protein, human; D003470:Culture Media; D053583:Interleukin-1beta; D007377:Interleukin-3; D015850:Interleukin-6; D007378:Interleukins; D063268:Sialic Acid Binding Ig-like Lectin 3