Expansion of neutrophil precursors and progenitors in suspension cultures of CD34+ cells enriched from human bone marrow.
SL Smith, JG Bender, PB Maples… - Experimental …, 1993 - europepmc.org
SL Smith, JG Bender, PB Maples, K Unverzagt, M Schilling, L Lum, S Williams, DE Van Epps
Experimental hematology, 1993•europepmc.orgThe growth and differentiation of selected bone marrow CD34+ cells stimulated with
hematopoietic growth factors in lipid cultures were evaluated to determine whether cell types
that may be useful for reducing the neutropenia associated with high-dose chemotherapy
(HDC) can be produced and quantitated in vitro. CD34+ cells enriched from bone marrow
were cultured for up to 5 weeks in interleukin-3 (IL-3), granulocyte-macrophage colony-
stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) with or …
hematopoietic growth factors in lipid cultures were evaluated to determine whether cell types
that may be useful for reducing the neutropenia associated with high-dose chemotherapy
(HDC) can be produced and quantitated in vitro. CD34+ cells enriched from bone marrow
were cultured for up to 5 weeks in interleukin-3 (IL-3), granulocyte-macrophage colony-
stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) with or …
The growth and differentiation of selected bone marrow CD34+ cells stimulated with hematopoietic growth factors in lipid cultures were evaluated to determine whether cell types that may be useful for reducing the neutropenia associated with high-dose chemotherapy (HDC) can be produced and quantitated in vitro. CD34+ cells enriched from bone marrow were cultured for up to 5 weeks in interleukin-3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) with or without stem cell factor (SCF)(also termed c-kit ligand). The mixture of IL-3, GM-CSF and G-CSF resulted in an 18-fold increase in cells after 10 to 12 days of culture and a 94-fold increase after 21 days. A 3-fold increase in colony-forming unit granulocyte-macrophage (CFU-GM) was observed after 10 days of culture. The addition of SCF during the first 10 days of culture further augmented the proliferation of cell numbers to 24-fold and colony-forming cells (CFC) to 8-fold after 10 days while cell numbers increased 130-fold after 21 days. Two-color flow cytometry defined phenotypes expressing CD11b and CD15 that represented maturation stages of neutrophils. Maturation of neutrophils in these cultures could be followed by the initial appearance after 3 to 7 days of a CD15+ CD11b-phenotype representing promyelocytes, which gave rise after 2 to 3 weeks to a CD15+ CD11b+ phenotype representing more mature neutrophil forms (metamyelocytes to segmented neutrophils). In contrast to normal neutrophil development, only a small fraction (10 to 15%) of the culture-derived neutrophils expressed CD16. These data define the kinetics and differentiation of neutrophils and neutrophil precursors from selected CD34+ cells in liquid cultures.
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