Hematopoietic Stem Cell Function in a Murine Model of Sickle Cell Disease
Figure 2
Stem cell number and oxidant state in Berkeley sickle mice. (a) Quantification of KSL progenitor cells in the bone marrow of control, hemizygous, and homozygous sickle mice showing a significant reduction in number in homozygous sickle mice. (b) Further examination of a phenotypically defined HSC population (KSL/CD150+/CD48−) also shows a reduction of HSC in sickle BM. (c) HSCs in the stem cell niche are known to be quiescent, and homozygous mice again show significant reduction in KSL cells in the G0 phase of the cell cycle possibly indicating the mobilization of sickle progenitor cells. (d) The in vitro colony-forming, high proliferative potential assay (HPP) of stem/progenitor cells further indicates a significant reduction in colony-forming cells in homozygous mice. (e) To examine the effect of oxidant stress on hematopoietic progenitors, we measured lipid peroxidation using the fluorescent indicator DHPE. A reduction in fluorescence indicates increased activity of hydroxyl radicals. (f) Further quantification of reactive oxygen species was shown in sickle mice by the increased production of DCF (all graphs are presented as mean ± SD, *, **, ***).
