The expansion of megakaryocyte progenitors from CD34+-enriched mobilized peripheral blood stem cells is inhibited by Flt3-L

J. Case, C. Hicks, A. Trickett, Y. L. Kwan, A. Manoharan

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


This study aimed to determine the optimal growth factor combination for expansion of megakaryocyte (Mk) progenitors with clonogenic potential from CD34+-enriched mobilized peripheral blood stem cells (PBSC). Mobilized PBSC were monocyte depleted and CD34+ enriched, then cultured with various combinations of interleukin-3 (IL-3), IL-6, IL-11, Flt3 ligand (Flt3-L), stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and erythropoietin (EPO), using a 2 7-3IV fractional factorial design. Expansion of Mk committed progenitors (CD41+) and primitive precursors (CD61 +CD34+) was determined using FACS and colony-forming assays. Amplification of Mk progenitor production was attributed to IL-3 (p < 0.002), SCF (p < 0.001), and GM-CSF (p < 0.05). Flt3-L inhibited the production of total CD61+ cells (p < 0.05), CD61 +CD34+ cells (p < 0.03), and total CD41a+ cells (p < 0.01). Addition of Flt3-L to the optimum growth factor combination of megakaryocyte growth and development factor (MGDF), SCF, IL-3, and GM-CSF caused the greatest increase in total nucleated cells but reduced Mk progenitor expansion. There was also a 20% reduction in Mk+ colonies from cells expanded in the presence of Flt3-L. Factorial analysis identified the optimal combination of growth factors required to expand Mk precursors with clonogenic potential. The addition of Flt3-L to the optimal combination of MGDF, SCF, IL-3, and GM-CSF reduced both the fold expansion of Mk progenitors and Mk colony numbers.

Original languageEnglish (US)
Pages (from-to)76-82
Number of pages7
JournalJournal of Interferon and Cytokine Research
Issue number2
StatePublished - Feb 1 2006
Externally publishedYes

ASJC Scopus subject areas

  • Immunology
  • Cell Biology
  • Virology

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