Research article

Hematopoietic progenitor cells and interleukin-stimulated endothelium: expansion and differentiation of myeloid precursors

Anja Moldenhauer¹ , Gesche Genter¹ , Andreas Lun² , Gürkan Bal¹ , Holger Kiesewetter¹ and Abdulgabar Salama¹

¹  Institute for Transfusion Medicine, Charité – Universitätsmedizin Berlin, Germany

²  Institute for Laboratory Medicine and Pathobiochemistry, Charité – Universitätsmedizin Berlin, Germany

author email corresponding author email

BMC Immunology 2008, 9:56doi:10.1186/1471-2172-9-56

Published:	1 October 2008

Abstract

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-1β, 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-1β 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-1β 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.

Conclusion

IL-1β and IL-3 stimulated endothelium induces proliferation and differentiation of myeloid precursors, while IL-6 treatment induced a benefit of HPC survival.