Abstract
Recent reports have identified CD34neg normal hematopoietic stem cells from several species based on their Hoechst 33342 dual emission staining profile. Side population (SP) cells exclude Hoechst 33342 (Hoechstlow) and are highly enriched in bone marrow repopulating activity in lethally irradiated mice (Goodell et al., 1997; Goodell et al., 1996). CD34neg stem cells were shown to engraft in NOD/scid mice,(Bhatia M and et al, 1998) and in fetal sheep (Zanjani et al. 1998). CML is thought to originate in hematopoietic stem cells. Much work has been done to characterize these cells and to identify a leukemia-free stem cell compartment. To determine if the SP compartment contained normal or leukemic cells in patients with CML, we sorted SP cells that were lineage negative using a cocktail of 14 different monoclonal antibodies as markers of lineage differentiation (SP Linneg) and performed fluorescence in situ hybridization (FISH) for the detection of Ph+ cells found in CML. Primary bone marrow samples were obtained from 18 patients with CML. In all 16 cases where SP cells were detected, a reduction in the percentage of Ph+ cells was found in SP cells as compared to non-SP cells. In 11/14 samples where sorting was done on SP Linneg compartment, the level of Ph+ cells in the SP Linneg population was at or below background positivity for the FISH assay (12%). The non-SP population of the same samples contained 20-55% Ph+ cells. Conclusion: Results suggest that in the majority of patients with CML, the SP Linneg cell compartment is free of Ph-positive cells detectable by FISH.KeywordsChronic Myeloid LeukemiaSide PopulationChronic Myeloid Leukemia PatientSide Population CellChronic Myeloid Leukemia SampleThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
Published Version
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