Generation of natural killer cells from long-term cultures of mouse bone marrow

Nat Immun. Jan-Feb 1993;12(1):1-16.

Abstract

The features of a mouse long-term bone marrow culture (LTBMC) system that produces natural killer (NK) cell activity are described. Over a 4-week period in the NK-LTBMC, cellularity dropped from approximately 2.5 x 10(7) to 8 x 10(5) cells/25-cm2 flask. About 3 x 10(5) of these cells were loosely adherent. The cultures at this time contained about one-third the spleen colony forming units, one-tenth the granulocyte macrophage colony forming units and about one-third the transplantable NK progenitor activity of fresh bone marrow (BM), and no detectable NK cell activity. In the 4-week NK-LTBMC, IL-2-responsive precursor cells appeared to be selectively maintained and gave an 8-fold higher activity after culture with human recombinant IL-2 (rIL-2) than did fresh BM. The addition of 50-5,000 IU/ml of rIL-2 resulted, after a minimal 3-day lag, in progressively increased cellularity for as long as 13 days. The percentage and staining intensity of NK-1.1+ cells increased with time after addition of rIL-2. CD3 epsilon + cells were occasionally seen and B220+ cells were present in low numbers at day 7 and slowly increased through day 13. The stroma was necessary for IL-2-dependent development of NK activity.

Publication types

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

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Bone Marrow / immunology*
  • Bone Marrow Cells
  • Cell Division
  • Cells, Cultured
  • Colony-Forming Units Assay
  • Cytotoxicity, Immunologic / immunology
  • Flow Cytometry
  • G(M1) Ganglioside / immunology
  • Immunophenotyping
  • Interleukin-2 / immunology
  • Killer Cells, Natural / cytology
  • Killer Cells, Natural / immunology*
  • Leukocyte Count
  • Longitudinal Studies
  • Mice
  • Mice, Inbred C57BL
  • Recombinant Proteins / immunology

Substances

  • Antibodies, Monoclonal
  • Interleukin-2
  • Recombinant Proteins
  • G(M1) Ganglioside
  • asialo GM1 ganglioside