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An Optimized Method for Isolating and Expanding Invariant Natural Killer T Cells from Mouse Spleen
Published on: October 29, 2015
Isolation and functional use of human NKT cells
Mark A Exley1, Brian Wilson, Steven P Balk
1Cancer Biology Program, Hematology-Oncology Division, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.
Current Protocols in Immunology
|September 4, 2010
Summary
This study provides detailed methods for isolating, expanding, and functionally analyzing human invariant natural killer T (iNKT) cells. These protocols enable researchers to better study these rare immune cells for various applications.
Area of Science:
- Immunology
- Cell Biology
Background:
- Invariant natural killer T (iNKT) cells are a rare population of immune cells with unique characteristics.
- These cells recognize the CD1d protein and express a conserved T cell receptor (TCR) alpha chain.
- Understanding iNKT cell biology is crucial for developing immunotherapies.
Purpose of the Study:
- To detail robust methods for the isolation, in vitro expansion, and functional characterization of human iNKT cells.
- To provide researchers with reproducible protocols for working with these specialized immune cells.
Main Methods:
- Multi-color Fluorescence-Activated Cell Sorting (FACS) analysis for identification and quantification.
- Cell sorting techniques for purification and isolation of iNKT cell clones.
- Secondary stimulation and rapid expansion protocols for iNKT cells.
- Functional assays for comprehensive analysis of iNKT cell activity.
Main Results:
- Established protocols for efficient isolation and purification of human iNKT cells from various samples.
- Demonstrated methods for in vitro expansion and generation of iNKT cell clones.
- Outlined techniques for functional characterization, enabling detailed study of iNKT cell responses.
Conclusions:
- The presented methods provide a comprehensive toolkit for researchers studying human iNKT cells.
- These protocols facilitate further investigation into the role of iNKT cells in immunity and disease.
- The ability to isolate, expand, and functionally analyze iNKT cells is key to advancing their therapeutic potential.
