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A Novel Feeder-free System for Mass Production of Murine Natural Killer Cells In Vitro
Published on: January 9, 2018
Isolation of murine natural killer cells
A Reichlin1, K Iizuka, W M Yokoyama
1Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, Missouri, USA.
Current Protocols in Immunology
|April 25, 2008
Summary
This protocol details isolating natural killer (NK) cells from mouse spleens. It uses antibody-based cell depletion and density gradients for high-purity NK cell isolation without antibody coating.
Area of Science:
- Immunology
- Cell Biology
Background:
- Natural killer (NK) cells are crucial for innate immunity.
- Efficient isolation of pure NK cells is essential for research and therapeutic applications.
Purpose of the Study:
- To describe a method for isolating highly purified natural killer (NK) cells from mouse spleen.
- To provide a protocol that avoids NK cell activation during isolation.
Main Methods:
- Negative selection using monoclonal antibodies (MAbs) to deplete contaminating cells.
- Complement lysis to eliminate unwanted cells.
- Density-gradient centrifugation to remove dead cells.
Main Results:
- Achieved a highly purified population of NK cells.
- The negative selection method prevents antibody-induced NK cell activation.
- Purity is confirmed by assessing cell surface phenotype.
Conclusions:
- This protocol provides an effective method for obtaining pure, non-activated NK cells from mouse spleen.
- The described technique is valuable for immunological studies requiring functional NK cells.
