Expansion of neutrophil precursors and progenitors in suspension cultures of CD34+ cells enriched from human bone marrow

S. L. Smith, J. G. Bender*, P. B. Maples, K. Unverzagt, M. Schilling, L. Lum, S. Williams, D. E. Van Epps

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

68 Scopus citations


The growth and differentiation of selected bone marrow CD34+ cells stimulated with hematopoietic growth factors in liquid 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.

Original languageEnglish (US)
Pages (from-to)870-877
Number of pages8
JournalExperimental Hematology
Issue number7
StatePublished - 1993


  • CD34
  • Colony forming cells
  • Colony-stimulating factors
  • Hematopoiesis
  • Neutrophil

ASJC Scopus subject areas

  • Molecular Biology
  • Hematology
  • Genetics
  • Cell Biology
  • Cancer Research


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