Enrichment of murine marrow cells for progenitors of multilineage hemopoietic colonies

Leukemia. 1992 Mar;6(3):193-8.


We attempted to develop a purification method for cell cycle-dormant murine multipotential progenitors that is simple and has a high recovery. Multilineage colony formation from mice that had been treated with 150 mg/kg 5-fluorouracil was assayed in cultures containing interleukin-3, interleukin-6, and erythropoietin. The cells with density 1.0631-1.0770 g/cm3 were approximately 40-fold enriched for multipotential progenitors. Negative immunomagnetic selection with a panel of lineage-specific monoclonal antibodies including anti-B220, anti-Gr-1, anti-Mac-1, anti-L3T4, and anti-Lyt-2 resulted in a cumulative 150-fold enrichment. When the density-separated and lineage-negative cells were further enriched by fluorescence-activated cell sorting with monoclonal antibody D7 (anti-Ly-6A/E), total enrichment averaged 800-fold and recovery was 17%. The method described here provides a relatively simple technique for the isolation of the multipotential progenitors and should be useful for the studies of regulation of hemopoiesis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Bone Marrow Cells*
  • Cell Count
  • Cell Cycle
  • Cell Separation
  • Cells, Cultured
  • Colony-Forming Units Assay
  • Culture Media
  • Erythropoietin / pharmacology
  • Female
  • Flow Cytometry
  • Fluorouracil / pharmacology
  • Hematopoietic Stem Cells / cytology*
  • Interleukin-3 / pharmacology
  • Interleukin-6 / pharmacology
  • Male
  • Mice
  • Mice, Inbred Strains


  • Antibodies, Monoclonal
  • Culture Media
  • Interleukin-3
  • Interleukin-6
  • Erythropoietin
  • Fluorouracil