title: Expansion of neutrophil precursors and progenitors in suspension cultures of CD34+ cells enriched from human bone marrow.
abstract: 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.
authors.affiliation: Immunotherapy Division, Baxter Healthcare Corp., Round Lake, IL 60073.
authors.forename: S L
authors.initials: SL
authors.lastname: Smith
authors.forename: J G
authors.initials: JG
authors.lastname: Bender
authors.forename: P B
authors.initials: PB
authors.lastname: Maples
authors.forename: K
authors.initials: K
authors.lastname: Unverzagt
authors.forename: M
authors.initials: M
authors.lastname: Schilling
authors.forename: L
authors.initials: L
authors.lastname: Lum
authors.forename: S
authors.initials: S
authors.lastname: Williams
authors.forename: D E
authors.initials: DE
authors.lastname: Van Epps
mesh_terms: D015703:Antigens, CD / Q000032:analysis*; D018952:Antigens, CD34; D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis; D001853:Bone Marrow / Q000276:immunology; D001854:Bone Marrow Cells; D018845:CD11 Antigens; D002448:Cell Adhesion; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D005434:Flow Cytometry; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D007377:Interleukin-3 / Q000494:pharmacology; D016256:Lewis X Antigen; D009504:Neutrophils / Q000166:cytology* / Q000276:immunology; D019089:Stem Cell Factor
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; D015214:Antigens, Differentiation, Myelomonocytic; D018845:CD11 Antigens; D016298:Hematopoietic Cell Growth Factors; D007377:Interleukin-3; D016256:Lewis X Antigen; D019089:Stem Cell Factor; D016179:Granulocyte Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor