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Murine neutrophils collected from subcutaneously implanted sponges
Abstract:
Polyvinyl sponges implanted subcutaneously in mice were used as a source of murine neutrophils (PMNs). Yield of PMNs from sponges averaged 6.3 X 10(5) PMNs per mouse and viability of sponge PMNs was 96.5%. These results were comparable to the yield and viability of PMNs harvested from peripheral blood. The chemotactic activity of sponge PMNs was significantly greater than the chemotactic activity of peripheral blood PMNs (chemotactic index of 4.78 +/- 1.66 in sponge PMNs versus 2.16 +/- 1.08 in peripheral blood PMNs, P less than 0.001). The difference in chemotactic activity was not attributable to hypotonic injury of PMNs nor the presence of soluble factors in sponge fluids. Phagocytic activity of sponge PMNs was comparable to the activity of blood PMNs (mean of 31.3 +/- 16.4% phagocytic cells for sponge PMNs versus 33.0 +/- 23.9% for blood PMNs). In addition, the number of fluorescent spheres ingested by sponge PMNs was not different (mean of 2.81 +/- 0.86 in sponge PMNs versus 2.65 +/- 0.68 spheres per cell for blood PMNs). These studies indicate that subcutaneously implanted sponges can be used as a source of functioning murine PMNs.
Insights
Subcutaneous polyvinyl sponges provide a rich source of murine neutrophils (PMNs). These sponge-derived PMNs exhibit enhanced chemotactic activity compared to peripheral blood PMNs, while maintaining comparable phagocytic function.
Area of Science:
- Immunology
- Cell Biology
Background:
- Murine neutrophils (PMNs) are crucial immune cells.
- Obtaining sufficient quantities of functional PMNs for research can be challenging.
Purpose of the Study:
- To evaluate subcutaneously implanted polyvinyl sponges as a novel source for murine neutrophils (PMNs).
- To compare the yield, viability, chemotactic, and phagocytic activities of PMNs isolated from sponges versus peripheral blood.
Main Methods:
- Polyvinyl sponges were implanted subcutaneously in mice.
- Murine neutrophils (PMNs) were harvested from sponges and peripheral blood.
- Cell yield, viability, chemotactic activity, and phagocytic activity were assessed and compared.
Main Results:
- Sponge implantation yielded an average of 6.3 X 10(5) PMNs per mouse with 96.5% viability.
- Sponge-derived PMNs demonstrated significantly greater chemotactic activity (chemotactic index 4.78) than peripheral blood PMNs (2.16).
- Phagocytic activity and particle uptake were comparable between sponge and blood PMNs.
Conclusions:
- Subcutaneously implanted polyvinyl sponges are an effective method for isolating functional murine neutrophils (PMNs).
- This technique provides a viable alternative for obtaining PMNs with enhanced chemotactic properties for immunological studies.