Self-generating density gradients of Percoll provide a simple and rapid method that consistently enriches natural killer cells

Stuart E. Ravnik, Sarah Gage, Sylvia B. Pollack

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

The separation or enrichment of natural killer (NK) cells from the heterogeneous cell populations in murine spleen or bone marrow is a vital step for the study of NK cells. We report in this study a simple and rapid method for the enrichment of NK cells from B cell-depleted spleen cells, using a self-generating density gradient of polyvinyl pyrrolidone-coated silica (Percoll). Nylon wool-passed spleen cells are suspended in Percoll that is isotonic and isosmotic with mouse blood at a density of 1.087 g/ml and ultracentrifuged at 30 000 × g for 10 min. This method consistently enriches for NK cell cytotoxic activity, in spleen cells of both unstimulated and interferon-stimulated mice, as measured in the chromium release assay. There is a concomitant enrichment for cells bearing the NK marker asialo GM-1 and depletion of L3T4 or Lyt-2-bearing T cells. In contrast to discontinuous, step-wise gradients, the self-generating Percoll gradient, which relies on the intrinsic property of Percoll to form a continuous density gradient, appears to provide the cells with a physiological environment both before and during the centrifugation step.

Original languageEnglish (US)
Pages (from-to)161-168
Number of pages8
JournalJournal of Immunological Methods
Volume110
Issue number2
DOIs
StatePublished - Jun 13 1988

Keywords

  • Cytotoxicity
  • Natural killer cell
  • Percoll
  • Self-generating gradient

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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