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CML and Stemness of Primitive CD34+ Cells

Autonomous multi-lineage differentiation in vitro of primitive CD34+ cells from patients with chronic myeloid leukemia

Abstract

Neoplastic CD34+ cells from chronic myeloid leukemia (CML) patients proliferate in vitro in the absence of serum or defined growth factors due to an autocrine mechanism involving IL-3 and G-CSF (Jiang et al. Proc Natl Acad Sci USA1999; 96: 12804). Detailed examination of the various cell types produced in such cultures has now demonstrated the rapid, factor-independent, generation of clonogenic progenitors for all lineages (granulocyte–macrophage, megakaryocyte and erythroid) with the additional appearance within 10 days of large numbers of mature granulocytes, macrophages, and megakaryocytes, as well as occasional erythroid cells. Inclusion of flt3-ligand, Steel factor, IL-3, IL-6, and G-CSF ± erythropoietin (EPO) in the cultures enhanced only slightly the output of mature cells (except for the erythroid population which was much larger when EPO was added). Analogous subpopulations of normal CD34+ cells produced similar numbers and types of cells but, as expected, only when growth factors were added. Thus primitive CD34+ CML cells proliferating autonomously in vitro recapitulate the full spectrum of differentiation responses of normal CD34+ cells stimulated by IL-3 and G-CSF. These findings point to a role of autocrine IL-3 and G-CSF in the similar multi-lineage expansion of differentiating CD34+ CML cells that occurs in vivo.

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Acknowledgements

The authors thank Dr Michael Barnett and the other members of the Division of Hematology of the BC Cancer Agency, the Vancouver Hospital and the University of British Columbia for assistance in procuring cells from normal donors and patients with CML and the associated clinical data. They also thank Terry Fox Laboratory staff in the Stem Cell Assay Service for assistance in initial cell processing, in the FACS facility for assistance in cell sorting, and Yvonne Yang for secretarial help. Cangene, Genentech, Immunex, Novartis, StemCell and P Lansdorp (Terry Fox Laboratory) generously provided gifts of cytokines and antibodies. This work was supported by a grant from the National Cancer Institute of Canada (NCIC) with funds from the Terry Fox Run. T Holyoake holds a United Kingdom Leukemia Research Fund Senior Lecturership, W Eisterer is an Austrian Government Scholarship Fellow (FWF, Erwin Schrodinger-Auslandsstipendium), M Berger was supported by a grant from the Association pour la Recherche Contre le Cancer (France). C Eaves is a Terry Fox Cancer Research Scientist of the NCIC.

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Jiang, X., Fujisaki, T., Nicolini, F. et al. Autonomous multi-lineage differentiation in vitro of primitive CD34+ cells from patients with chronic myeloid leukemia. Leukemia 14, 1112–1121 (2000). https://doi.org/10.1038/sj.leu.2401752

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