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Generation of a conditionally immortalized myeloid progenitor cell line requiring the presence of both interleukin-3 and stem cell factor to survive and proliferate.
Lee C, Evans C, Spooncer E, Pierce A, Mottram R, Whetton A
Br J Haematol. 2003;122( 6):985-95.
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Abstract
The H-2Kappab temperature-sensitive (ts) A58 transgenic (Immorto) mouse has been used previously to generate conditionally immortalized cells from a number of tissues. The present study aimed to investigate characteristics of primitive myeloid precursor cells derived from H-2Kappab-tsA58 bone marrow. Cell populations were enriched for granulocyte/macrophage progenitors by centrifugal elutriation, and were cultured in the presence and absence of cytokines at the permissive and restrictive temperatures for the A58 oncogene. Cells derived from H-2Kappab-tsA58 mice required both A58 activation and the growth factors, stem cell factor (SCF) and interleukin-3 (IL-3), for long-term cell survival and growth; cells were maintained for > 300 d in culture under these conditions. IL-3- and SCF-dependent clonal cell lines were derived with a phenotype (lin-, Sca-1+, CD34+, ER-MP 58+, ER-MP 12+, ER-MP 20-) characteristic of primitive myeloid progenitors. These cells differentiated on addition of granulocyte/macrophage colony-stimulating factor (GM-CSF) or macrophage colony-stimulating factor (M-CSF) and acquired mature cell morphology with some upregulation of differentiation markers. In conclusion, the A58 oncogene can immortalize haemopoietic progenitor cells. These cells require two cytokines for growth, IL-3 and SCF; as such, they constitute a useful resource for the study of synergistic interactions between growth factors. The ability to develop monocytic cell characteristics also permits the investigation of cytokine-mediated early haemopoietic progenitor cell development.
Keyword(s)
Animals; Cell Line; Cells, Cultured; Colony-Forming Units Assay; Mice; Mice, Transgenic; Research Support, Non-U.S. Gov't; cytology: Bone Marrow Cells; cytology: Hematopoietic Stem Cells; cytology: Stromal Cells; drug effects: Cell Differentiation; drug effects: Cell Division; genetics: H-2 Antigens; pharmacology: Growth Substances; pharmacology: Interleukin-3; pharmacology: Recombinant Proteins; pharmacology: Stem Cell Factor