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Isolating Lymphocytes from the Mouse Small Intestinal Immune System
Published on: February 28, 2018
Isolation of rat intestinal lymph DC
Simon Milling1, Gordon MacPherson
1Glasgow Biomedical Research Centre, University of Glasgow, Glasgow, UK.
Methods in Molecular Biology (Clifton, N.J.)
|November 27, 2009
Summary
Researchers developed a new method to collect intestinal lymph dendritic cells (ilDCs) from rats without causing maturation. This technique preserves the natural state of these crucial immune cells for further study.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) constantly migrate from the intestine to mesenteric lymph nodes.
- Intestinal lymph DCs (ilDCs) are vital for initiating immune responses and maintaining oral tolerance.
- ilDCs are sensitive to environmental changes, leading to maturation during extraction.
Purpose of the Study:
- To develop and refine a method for collecting and purifying rat ilDC subsets.
- To prevent DC maturation during the isolation process.
Main Methods:
- Surgical removal of mesenteric lymph nodes followed by thoracic duct cannulation.
- Collection of ilDCs on ice immediately after exiting lymph vessels.
- Enrichment using magnetic beads and purification via flow cytometric sorting.
- Refinements to minimize animal restraint.
Main Results:
- A refined protocol for isolating ilDCs without inducing maturation was established.
- The method allows for the collection of distinct ilDC subsets.
Conclusions:
- The described method provides a reliable way to obtain immature ilDCs.
- This technique is crucial for studying the in vivo functions of ilDCs in immunity and tolerance.
