Isolation and analysis of primitive hemopoietic progenitor cells on the basis of differential expression of Qa-m7 antigen

J Immunol. 1986 May 1;136(9):3219-24.

Abstract

In the presence of the hemopoietic growth factor CSF-1, the later committed cells of the macrophage lineage can be detected by their ability to form small colonies in clonal agar culture (CFCCSF-1). Synergistic factors have been described that in combination with CSF-1 stimulate developmentally early hemopoietic progenitor cells of high proliferative potential (HPP-CFC). By using a monoclonal antibody to the Qa-m7 antigenic determinant, we investigated and compared the expression of Qa-m7 on CFCCSF-1 and on HPP-CFC of two types that grow in response to either 1) CSF-1 plus synergistic factor from human placenta-conditioned medium (HPP-CFCHplac+CSF-1) or 2) CSF-1 plus synergistic factor from conditioned medium of the WEHI-3 myelomonocytic cell line (HPP-CFCW+CSF-1). We have shown that HPP-CFC of both types express relatively more Qa-m7 antigen than CFCCSF-1 and can be separated and enriched on this basis by discontinuous buoyant density centrifugation and fluorescence-activated cell sorting of normal bone marrow. Significant enrichments of HPP-CFCHPlac+CSF-1 (43.5-fold) and HPP-CFCW+CSF-1 (28.8-fold) have been achieved with cloning efficiencies of HPP-CFC in the most enriched fractions reaching 4 to 5%. These results clearly illustrate the fact that there are populations of progenitor cells from normal, unperturbed bone marrow that strictly require a combination of two hemopoietic growth factors (CSF-1 plus synergistic factor) in order to be detected.

Publication types

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

MeSH terms

  • Animals
  • Antigens, Surface / analysis*
  • Antigens, Surface / immunology
  • Cell Differentiation
  • Cell Separation* / methods
  • Centrifugation, Density Gradient
  • Colony-Forming Units Assay
  • Cytotoxicity Tests, Immunologic
  • Flow Cytometry
  • Hematopoietic Stem Cells / classification*
  • Hematopoietic Stem Cells / immunology
  • Histocompatibility Antigens / analysis*
  • Histocompatibility Antigens / immunology
  • Histocompatibility Antigens Class I*
  • Lymphocyte Activation
  • Mice
  • Mice, Inbred Strains

Substances

  • Antigens, Surface
  • Histocompatibility Antigens
  • Histocompatibility Antigens Class I
  • Q surface antigens