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

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

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

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 7-3 IV 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
Volume26
Issue number2
DOIs
StatePublished - Feb 2006
Externally publishedYes

Fingerprint

Megakaryocyte Progenitor Cells
Stem Cell Factor
Interleukin-3
Granulocyte-Macrophage Colony-Stimulating Factor
Megakaryocytes
Thrombopoietin
Intercellular Signaling Peptides and Proteins
Interleukin-11
Erythropoietin
Monocytes
Interleukin-6
Peripheral Blood Stem Cells
flt3 ligand protein

ASJC Scopus subject areas

  • Immunology
  • Virology
  • Cell Biology

Cite this

The expansion of megakaryocyte progenitors from CD34+-enriched mobilized peripheral blood stem cells is inhibited by Flt3-L. / Case, Jamie; Hicks, C.; Trickett, A.; Kwan, Y. L.; Manoharan, A.

In: Journal of Interferon and Cytokine Research, Vol. 26, No. 2, 02.2006, p. 76-82.

Research output: Contribution to journalArticle

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abstract = "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 7-3 IV 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.",
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