Detection of exogenous DNA uptake by murine dendritic cells

• Published in: STAR protocols Vol. 3; no. 3; p. 101464
• Main Authors: Celias, Daiana P., de Mingo Pulido, Álvaro, Ruffell, Brian
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16.09.2022; Elsevier
• Subjects: Animals; Cancer; Cell Biology; Cell isolation; Dendritic Cells; DNA; Flow Cytometry/Mass Cytometry; Immunology; Mice; Microscopy; Molecular/Chemical Probes; Protocol; Immunology; Microscopy; Flow Cytometry/Mass Cytometry; Molecular/Chemical Probes; Cell isolation; Cancer; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101464

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). [Display omitted] •In vitro assay to measure exogenous DNA uptake by primary murine dendritic cells•Designed to mimic conditions of high cell death such as those within tumors•DNA uptake occurs spontaneously without the use of transfection reagents•Useful for studying DNA uptake and searching for activators or inhibitors Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.