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Updated: Dec 3, 2025
Bone Marrow-derived Macrophage Production
Published on: November 22, 2013
Bacterial Synchronized Transfer Assays in Bone Marrow Derived Macrophages
Sedelia R Dominguez1, Tom Kawula1, Shaun P Steele1
1Paul G Allen School of Global Animal Health, Washington State University, Pullman, United States.
Abstract:
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.
Insights
Merocytophagy, a cell-eating process, was studied using a novel synchronized transfer assay. This method enables high-purity isolation of recipient cells for detailed analysis of microbial and cytosolic material transfer.
Area of Science:
- Cell Biology
- Microbiology
- Immunology
Background:
- Merocytophagy involves partial cell eating upon cell-cell contact, transferring microbes and cytosolic material.
- Existing methods for studying cell-cell contact dependent transfer lack recipient cell isolation, hindering detailed analysis.
- Analyzing recipient cells and bacterial burden within them is challenging with current co-incubation assays.
Purpose of the Study:
- To develop and present a synchronized transfer assay for studying merocytophagy.
- To enable high-purity isolation of recipient cells following transfer events.
- To facilitate investigation of cell-cell contact dependent transfer mechanisms and post-merocytophagy events.
Main Methods:
- Establishment of a synchronized transfer assay.
- Co-incubation of differently labeled cells to assess transfer events.
- Isolation of recipient cells from donor cells at high purity post-transfer.
- Application of the assay in the context of bacterial infections and cytosolic cellular staining.
Main Results:
- The synchronized transfer assay allows for recipient cell isolation with high purity.
- The assay facilitates detailed analysis of merocytophagy events.
- Bacterial burden and cytosolic material transfer into recipient cells can be accurately assessed.
Conclusions:
- The developed synchronized transfer assay is a valuable tool for investigating merocytophagy.
- This protocol simplifies the study of cell-cell contact dependent transfer mechanisms.
- The assay enables deeper understanding of the events following merocytophagy, particularly in microbial infections.
