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Related Experiment Videos

A multi-step isolation scheme for obtaining CD16+ human natural killer cells.

L M Cosentino1, M K Cathcart

  • 1Department of Immunology and Cancer, Cleveland Clinic Foundation, OH.

Journal of Immunological Methods
|November 5, 1987
PubMed
Summary

A new multi-step method efficiently isolates CD16+ human natural killer (NK) cells. This enriched NK cell population shows enhanced cytotoxicity and higher recovery rates, facilitating NK cell research.

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Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Natural killer (NK) cells are crucial for innate immunity.
  • Efficient isolation of pure NK cell populations is vital for research.
  • Existing methods like Percoll density gradient centrifugation have limitations.

Purpose of the Study:

  • To develop and validate a novel multi-step protocol for isolating enriched CD16+ human NK cells.
  • To compare the efficacy of this new method with the established Percoll density gradient technique.
  • To assess the functional capacity and recovery yield of NK cells isolated by the new protocol.

Main Methods:

  • A multi-step negative selection protocol involving nylon wool incubation, sheep red blood cell (SRBC) rosetting, and panning.
  • Incubation of peripheral blood mononuclear cells (MNCs) with nylon wool.

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  • Rosetting non-adherent cells with SRBCs and panning to remove CD3+ cells.

    • Main Results:

    • The multi-step protocol yielded 70-80% CD16+ NK cells with minimal contamination.
    • Compared to Percoll density gradients, the new method provided higher CD16+ cell percentage, greater cytotoxicity, and fewer contaminating monocytes/macrophages.
    • Recovery of CD16+ cells was significantly higher (30%) with the multi-step method versus Percoll (7%).

    Conclusions:

    • The described multi-step isolation scheme consistently yields a high percentage of CD16+ NK cells.
    • This method offers superior efficiency and purity compared to Percoll density gradient centrifugation.
    • The enriched NK cells demonstrate enhanced cytotoxic activity and facilitate studies on NK cell-mediated cytotoxicity and cytotoxins.