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Chemotaxis as a preparative technique for human polymorphonuclear leukocytes.
The Journal of Laboratory and Clinical Medicine
|August 1, 1975
Summary
This study developed a method to isolate large numbers of human peripheral blood phagocytic cells, primarily polymorphonuclear leukocytes, using chemotaxis. This technique yields highly pure, biologically active cells for research.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Phagocytic cells, particularly polymorphonuclear leukocytes (PMNs), are crucial for innate immunity.
- Efficient isolation of pure, viable PMNs from human peripheral blood is essential for immunological studies.
- Existing methods may have limitations in yield, purity, or biological activity.
Purpose of the Study:
- To develop and optimize a method for large-scale isolation of human peripheral blood phagocytic cells.
- To utilize chemotaxis as the primary principle for cell separation.
- To obtain highly pure and biologically active phagocytic cell preparations.
Main Methods:
- Utilized an enlarged Boyden chemotactic chamber for cell isolation.
- Collected cells migrating through a Millipore membrane towards chemotactic factors.
- Tested various chemotactic factors including human serum and casein, with a combination proving most effective.
Main Results:
- Achieved yields of up to 24.8 x 10^6 cells from 12 c.c. of peripheral blood.
- Isolated cell populations were >95% polymorphonuclear leukocytes, with monocytes as the remainder.
- Preparations were virtually free of erythrocytes and lymphocytes, indicating high purity.
Conclusions:
- Chemotaxis-based isolation is an effective method for obtaining large quantities of pure human phagocytic cells.
- The isolated cells are biologically active, as confirmed by the chemotactic isolation requirement.
- This technique offers advantages over sedimentation-based methods, providing superior purity and cell viability.