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Isolation of Myeloid Dendritic Cells and Epithelial Cells from Human Thymus
Published on: September 19, 2013
A novel method for isolating dendritic cells from human bronchoalveolar lavage fluid
B Ten Berge1, F Muskens, A Kleinjan
1Department of Pulmonary Medicine, Erasmus MC, Dr. Molewaterplein 50, 3015 GE Rotterdam, The Netherlands.
Journal of Immunological Methods
|October 7, 2009
Summary
Researchers developed a new method to isolate dendritic cells (DCs) from bronchoalveolar lavage fluid. This technique yields functional myeloid DCs (mDCs) and plasmacytoid DCs (pDCs) for pulmonary disease research.
Area of Science:
- Immunology
- Pulmonary Medicine
Background:
- Dendritic cells (DCs) are crucial for immune responses and implicated in lung diseases.
- Current methods for isolating lung DCs are invasive or alter cell function.
- Bronchoalveolar lavage (BAL) offers a minimally invasive source of local DCs.
Purpose of the Study:
- To develop and validate a novel method for isolating functional dendritic cells (DCs) from bronchoalveolar lavage (BAL) fluid.
- To enable detailed studies of DC function in pulmonary diseases.
Main Methods:
- Developed a novel sorting strategy using fluorescence-activated cell sorting (FACS ARIA) on BAL cells.
- Identified myeloid DCs (mDCs) as lineage-negative, CD11c+, HLA-DR+ and plasmacytoid DCs (pDCs) as CD11c-, CD123+.
- Assessed purity, morphology, and function (mixed leukocyte reaction, T cell interaction) of sorted DCs.
Main Results:
- Successfully isolated high-purity mDCs and pDCs from BAL fluid.
- Sorted mDCs demonstrated characteristic morphology and induced T cell proliferation and differentiation.
- DC function was preserved after freezing and thawing.
Conclusions:
- A novel FACS-based sorting strategy enables isolation of functional mDCs from BAL fluid.
- This method provides a valuable tool for studying DCs in the context of pulmonary disease.
