Rhodamine123 reveals heterogeneity within murine Lin-, Sca-1+ hemopoietic stem cells.
Murine bone marrow Lin-, Ly6A/E+ cells have been fractionated on the basis of rhodamine123 retention into Rh123med/hi and Rh123lo subpopulations. These populations have different responses to hemopoietic growth factors with respect to in vitro colony formation. Cells from either fraction were not stimulated by only granulocyte-colony-stimulating factor (G-CSF), granulocyte/macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), interleukins 1 and 6 (IL-1 and -6), or leukemia inhibitory factor (LIF) alone. The Rh123med/hi, but not the Rh123lo fraction, contained cells that could be stimulated by either stem cell factor (SCF) or IL-3 alone. When combinations of growth factors were added, the Rh123med/hi fraction produced more colonies, and responded to a wider range of factor combinations than the Rh123lo population. When tested in vivo, both populations contained no detectable day 8 colony-forming unit-spleen (CFU-S), and similar frequencies of day 13 CFU-S. When transplanted into lethally irradiated recipients (100 cells/recipient), significant numbers of donor cells (67-73%) were found in the peripheral blood of Rh123lo recipients. Both myeloid and lymphoid cells were of donor origin. By comparison, the Rh123med/hi population produced recipients with 1-2% donor cells in peripheral blood, the majority of which were lymphoid.
Li
C L
CL
Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
Johnson
G R
GR
D000818:Animals; D000950:Antigens, Ly / Q000032:analysis*; D001854:Bone Marrow Cells; D002469:Cell Separation; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D006131:Growth Inhibitors / Q000494:pharmacology; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D007375:Interleukin-1 / Q000494:pharmacology; D007377:Interleukin-3 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000494:pharmacology; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D008565:Membrane Proteins / Q000032:analysis*; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D010641:Phenotype; D020112:Rhodamine 123; D012235:Rhodamines; D019089:Stem Cell Factor
D000950:Antigens, Ly; D005456:Fluorescent Dyes; D006131:Growth Inhibitors; D016298:Hematopoietic Cell Growth Factors; D007375:Interleukin-1; D007377:Interleukin-3; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; C074650:Ly6a protein, mouse; D008222:Lymphokines; D008565:Membrane Proteins; D012235:Rhodamines; D019089:Stem Cell Factor; D016179:Granulocyte Colony-Stimulating Factor; D020112:Rhodamine 123; D016173:Macrophage Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor