Purification of murine bone-marrow-derived granulocyte-macrophage colony-forming cells

D. E. Williams, J. E. Straneva, R. N. Shen, H. E. Broxmeyer

Research output: Contribution to journalArticle

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Abstract

Previous attempts to purify progenitor cells that form colonies and clusters of granulocytes and/or macrophages (CFU-GM) from adult murine bone marrow have had limited success because of the paucity of these cells. In the present paper we report studies with a rapid, reproducible method involving pretreatment of mice, three days prior to sacrifice, with 200 mg/kg of Cytoxan (cyclophosphamide), density separation on Ficoll-Hypaque, and counterflow centrifugal elutriation, that yielded highly enriched populations of CFU-GM. The peak CFU-GM-containing fraction (FR-28) eluted at a flow rate of 28 ml/min and contained very little contamination by other in vitro colony-forming cells (BFU-E, CFU-GEMM, CFU-MK). FR-28 contained 0.54% ± 0.30% (16 experiments) of the unfractionated post-Cytoxan bone marrow nucleated cells and lacked significant contamination by lymphocytes and monocytes. The mean CFU-GM cloning efficiency of FR-28 was 44% ± 9% in agar (11 experiments) and 75% ± 10% in agarose (nine experiments). CFU-GM from FR-28 demonstrated linear plating characteristics even at very low cell density (25 cells), and formed colonies and clusters of granulocytes, macrophages, or both in the same proportions as did unfractionated post-Cytoxan or untreated bone marrow. Approximately 10% (assuming a seeding efficiency of 10%) of FR-28 cells were in vivo spleen colony-forming cells (CFU-S) measured at day 12. These results represent the highest degree of purity (up to 94%) of CFU-GM thus far reported and should prove useful in studies of this cell population.

Original languageEnglish
Pages (from-to)243-250
Number of pages8
JournalExperimental Hematology
Volume15
Issue number3
StatePublished - 1987

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Granulocyte-Macrophage Progenitor Cells
Granulocytes
Macrophages
Cyclophosphamide
Myeloid Progenitor Cells
Bone Marrow
Diatrizoate
Ficoll
Erythroid Precursor Cells
Bone Marrow Cells
Sepharose
Population
Agar
Organism Cloning
Monocytes
Stem Cells
Spleen
Cell Count
Lymphocytes

ASJC Scopus subject areas

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

Cite this

Purification of murine bone-marrow-derived granulocyte-macrophage colony-forming cells. / Williams, D. E.; Straneva, J. E.; Shen, R. N.; Broxmeyer, H. E.

In: Experimental Hematology, Vol. 15, No. 3, 01.01.1987, p. 243-250.

Research output: Contribution to journalArticle

Williams, D. E. ; Straneva, J. E. ; Shen, R. N. ; Broxmeyer, H. E. / Purification of murine bone-marrow-derived granulocyte-macrophage colony-forming cells. In: Experimental Hematology. 1987 ; Vol. 15, No. 3. pp. 243-250.
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