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Isolation of Mouse Lung Dendritic Cells
Published on: November 22, 2011
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Isolation of Conventional Dendritic Cells from Mouse Lungs
Lianne van de Laar1,2, Martin Guilliams3,4, Simon Tavernier3,5
1Laboratory of Immunoregulation, VIB Inflammation Research Center, Ghent University, Technologiepark 927, 9052, Ghent, Belgium. lianne.vandelaar@irc.ugent.be.
Methods in Molecular Biology (Clifton, N.J.)
|May 5, 2016
Summary
This study details methods for isolating lung dendritic cells (cDCs), crucial for understanding immune responses. The optimal isolation technique for these CD103(+) and CD11b(+) cDCs depends on experimental needs and cell quantity.
Area of Science:
- Immunology
- Cell Biology
- Respiratory Medicine
Background:
- The lung immune system constantly encounters environmental threats like pathogens and allergens.
- Lung dendritic cells (cDCs) are key immune sentinels, sampling antigens from the airway lumen.
- Distinct subsets of lung cDCs, including CD103(+) and CD11b(+) cells, possess unique immunological functions.
Purpose of the Study:
- To describe and compare protocols for the high-quality ex vivo isolation of lung CD103(+) and CD11b(+) conventional dendritic cells (cDCs).
- To provide guidance on selecting the optimal isolation method based on experimental requirements and desired cell yield.
Main Methods:
- Comparison of various cell isolation techniques for lung cDCs.
- Flow cytometry-assisted cell sorting (FACS) for low cell number requirements.
- Positive selection of CD11c(+) cells followed by FACS for high cell numbers intended for lysis or fixation.
- Gradient selection combined with sorting for isolating large quantities of cDCs for functional assays.
Main Results:
- Different isolation methods yield varying cell numbers and purities suitable for distinct experimental applications.
- Flow cytometry-assisted cell sorting (FACS) is effective for obtaining small numbers of specific cDC subsets.
- Positive selection followed by FACS is advantageous for experiments requiring large cell numbers that will be lysed or fixed.
- Gradient selection and subsequent sorting provide optimal yields for functional assays demanding substantial cell populations.
Conclusions:
- The choice of ex vivo isolation protocol for lung cDCs significantly impacts experimental outcomes.
- Tailoring the isolation method to specific experimental needs, such as cell number and downstream application, is critical for advancing research on lung immune responses.
