Related Experiment Video
Updated: Jul 9, 2025
Author Spotlight: Studying Macrophage-Epithelial Cell Interactions in Salivary Gland Regeneration After Injury
Published on: November 17, 2023
Interrogating Cell-Cell Interactions in the Salivary Gland via Ex Vivo Live Cell Imaging
Sonia Elder1, Justyna Cholewa-Waclaw1, Elaine Emmerson2
1The Centre for Regenerative Medicine, Institute for Regeneration and Repair, The University of Edinburgh.
Insights
This study presents a new method for live imaging of salivary gland tissue, enabling real-time observation of macrophage behavior during regeneration after injury. This advances understanding of salivary gland repair mechanisms.
Area of Science:
- Regenerative Medicine
- Cell Biology
- Immunology
Background:
- Salivary gland regeneration involves complex cell interactions, with macrophages playing a key role.
- Current understanding relies on static tissue analysis, limiting real-time insights into dynamic processes.
- Investigating real-time macrophage behavior is crucial for understanding salivary gland repair.
Purpose of the Study:
- To develop and validate a protocol for live imaging of ex vivo salivary gland tissue.
- To enable real-time observation of cell migration and interactions, particularly macrophages, during regeneration.
- To provide a tool for studying salivary gland repair dynamics post-injury.
Main Methods:
- Mouse submandibular salivary gland tissue slices cultured ex vivo at an air-liquid interface.
- Induction of injury (e.g., radiation) to trigger regenerative response.
- Live imaging of endogenously labeled or antibody-stained cells using confocal microscopy over 12 hours at 15-minute intervals.
Main Results:
- Successful establishment of a protocol for live imaging of salivary gland explants.
- Generation of time-lapse movies capturing cell migration and interactions.
- Extraction of cell behavior parameters from imaging data.
Conclusions:
- The developed ex vivo culture and live imaging method allows real-time investigation of macrophage roles in salivary gland regeneration.
- This approach overcomes limitations of static analyses, offering dynamic insights into cellular mechanisms of repair.
- Facilitates deeper understanding of salivary gland injury response and potential therapeutic targets.
Abstract:
Salivary gland regeneration is a complex process involving intricate interactions among various cell types. Recent studies have shed light on the pivotal role played by macrophages in driving the regenerative response. However, our understanding of this critical role has primarily relied on static views obtained from fixed tissue biopsies. To overcome this limitation and gain insights into these interactions in real time, this study outlines a comprehensive protocol for culturing salivary gland tissue ex vivo and capturing live images of cell migration. The protocol involves several key steps: First, mouse submandibular salivary gland tissue is carefully sliced using a vibratome and then cultured at an air-liquid interface. These slices can be intentionally injured, for instance, through exposure to radiation, to induce cellular damage and trigger the regenerative response. To track specific cells of interest, they can be endogenously labeled, such as by utilizing salivary gland tissue collected from genetically modified mice where a particular protein is marked with green fluorescent protein (GFP). Alternatively, fluorescently-conjugated antibodies can be employed to stain cells expressing specific cell surface markers of interest. Once prepared, the salivary gland slices are subjected to live imaging using a high-content confocal imaging system over a duration of 12 h, with images captured at 15 min intervals. The resulting images are then compiled to create a movie, which can subsequently be analyzed to extract valuable cell behavior parameters. This innovative method provides researchers with a powerful tool to investigate and better understand macrophage interactions within the salivary gland following injury, thereby advancing our knowledge of the regenerative processes at play in this dynamic biological context.
