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Updated: Sep 7, 2025
Assessing the Development of Murine Plasmacytoid Dendritic Cells in Peyer's Patches Using Adoptive Transfer of Hematopoietic Progenitors
Published on: March 17, 2014
Detection of exogenous DNA uptake by murine dendritic cells
Daiana P Celias1, Álvaro de Mingo Pulido2, Brian Ruffell3
1Department of Immunology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL 33612, USA.
Abstract:
This protocol has been developed to measure exogenous DNA uptake by murine dendritic cells (DCs) using supernatant containing cellular debris, which allows for DNA uptake in the absence of transfection reagents. Inhibitors or antibodies that alter the process can be added, and either flow cytometry or fluorescent microscopy can be used to measure DNA uptake. This is intended to mimic the exposure of DCs to dying cells in the tumor microenvironment or other pathological conditions of high cellular death. For complete details on the use and execution of this protocol, please refer to de Mingo Pulido et al. (2021).
Insights
This study presents a new method to measure how dendritic cells (DCs) take up external DNA from dead cells without needing transfection. This technique helps understand immune cell interactions in disease.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) play a crucial role in immune responses.
- Exposure to exogenous DNA from dying cells is common in pathological conditions like tumors.
- Understanding DNA uptake by DCs is vital for immune modulation.
Purpose of the Study:
- To develop and present a protocol for measuring exogenous DNA uptake by murine dendritic cells (DCs).
- To enable DNA uptake measurement without transfection reagents, mimicking in vivo conditions.
- To provide a method for studying inhibitors or antibodies affecting DNA uptake.
Main Methods:
- Utilizes supernatant containing cellular debris for DNA uptake.
- Allows for the addition of inhibitors or antibodies to study their effects.
- Employs flow cytometry or fluorescent microscopy for quantitative measurement of DNA uptake.
Main Results:
- Successfully established a protocol for measuring exogenous DNA uptake by DCs.
- Demonstrated DNA uptake occurs naturally from cellular debris.
- Validated the use of flow cytometry and microscopy for assessment.
Conclusions:
- The developed protocol offers a robust method to study exogenous DNA uptake by dendritic cells.
- This technique is valuable for investigating DC behavior in the tumor microenvironment and other disease states.
- Provides a foundation for developing targeted immunotherapies.
