Bacterial Synchronized Transfer Assays in Bone Marrow Derived Macrophages

Merocytophagy ("mero", Greek for partial; "cytophagy" for cell eating) is a process by which cells acquire microbes and cytosolic material through phagocytosis of a small portion of neighboring cells upon cell-cell contact. Cell-cell contact dependent transfer events can be asses...

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Bibliographic Details
Published in:Bio-protocol 2019-11, Vol.9 (22)
Main Authors: Dominguez, Sedelia R, Kawula, Tom, Steele, Shaun P
Format: Article
Language:English
Subjects:
Biology
Methods
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Summary:Merocytophagy ("mero", Greek for partial; "cytophagy" for cell eating) is a process by which cells acquire microbes and cytosolic material through phagocytosis of a small portion of neighboring cells upon cell-cell contact. Cell-cell contact dependent transfer events can be assessed through co-incubation of differently labeled cells. With these assays, it is difficult to analyze the recipient cells by microscopy or bacterial burden within only recipient cells. Therefore, we established a synchronized transfer assay that allows for recipient cells to be isolated from donor cells following transfer events at a high purity. Here, we present this assay in context of bacterial infections and cytosolic cellular staining. With this protocol, mechanisms of cell-cell contact dependent transfer events and the events following merocytophagy can easily be investigated.
ISSN:2331-8325
2331-8325
DOI:10.21769/bioprotoc.3437