In this protocol, Creative Biolabs outlines a comprehensive method for the isolation and identification of hamster macrophages, enabling researchers to delve into their unique biological functions and interactions. Through a combination of meticulous materials, precise methods, and insightful notes, this protocol is tailored to researchers seeking a burst of knowledge in the field.

Materials

• Hamster tissue samples: Obtain fresh tissue samples, preferably from spleen or lung, from euthanized hamsters.
• Sterile phosphate-buffered saline (PBS)
• Dulbecco's modified eagle medium (DMEM)
• Fetal bovine serum (FBS)
• Antibiotics
• Collagenase solution
• Red blood cell lysis buffer
• Fluorescent antibodies

Methods

Fig.1 Methods for isolation and identification of hamster macrophages. (Creative Biolabs Original)

1. Isolation of Hamster Macrophages
   Euthanize the hamster using appropriate methods and ensure adherence to ethical guidelines. Dissect and harvest peritoneal macrophages using aseptic techniques. Rinse the peritoneal cavity with sterile PBS and collect the lavage fluid. Centrifuge the lavage fluid and isolate peritoneal cells using lymphocyte separation medium. Wash the cells with sterile PBS and resuspend in medium supplemented with FBS and antibiotics.
2. Culture and Maintenance
   Plate the isolated macrophages in cell culture-grade plasticware. Incubate the cells at 37°C in a humidified atmosphere containing 5% CO2. Regularly change the medium to maintain cell viability and health.
3. Identification of Macrophages
   Perform cell staining with specific markers like F4/80 for macrophage identification. Use flow cytometry or immunofluorescence to analyze cell surface markers and confirm macrophage identity. Assess phagocytic activity by incubating macrophages with labeled particles and analyzing uptake.

Notes

• Proper animal handling and ethical considerations are essential throughout the protocol.
• Monitor cell purity through flow cytometry; adjust isolation methods if necessary.
• Choose antibodies based on hamster macrophage markers, considering cross-reactivity.
• Morphological assessment provides insights into cell integrity and potential activation states.
• Consider cytokine stimulation for functional studies, enhancing the burstiness of macrophage response.

In conclusion, this comprehensive protocol elucidates the intricate steps involved in isolating and identifying hamster macrophages. Creative Biolabs combines meticulous techniques with cutting-edge methods to help researchers can study macrophages in depth. For more species of macrophage isolation and identification, contact us.