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Influence of initial medium content (pH, serum and Ca++ concentration) on leukocyte migration test
1Department of Histology, Jagiellonian University, Collegium Medicum, Faculty of Medicine.
Abstract:
The aim of the study was to check to what extent inhibition of leukocyte migration in capillary tubes depends on alteration in pH, serum content and Ca++ concentration in medium in which the test is started. We found that relatively wide variations of pH (between 6.4 and 8.2) did not influence the final result. The best conditions for leukocyte migration were created by addition of 5% calf foetal serum, whereas a contamination of medium with autologous serum made a proper interpretation of the test difficult because of a resulting dispersion of cells. Neither removal nor addition of Ca++ ions had a significant effect on migration of leukocytes, unless combined with a simultaneous presence of calcium ionophore A 23187 in 0.5 microM concentration.
Insights
Leukocyte migration is not affected by pH or calcium levels. Calf serum enhances migration, while autologous serum hinders interpretation of leukocyte migration tests.
Area of Science:
- Immunology
- Cell Biology
Background:
- Leukocyte migration is a critical process in inflammatory responses.
- Understanding factors influencing leukocyte migration is essential for diagnostic assays.
Purpose of the Study:
- To investigate the impact of pH, serum content, and calcium ion concentration on leukocyte migration in vitro.
- To determine optimal conditions for leukocyte migration assays.
Main Methods:
- Leukocyte migration inhibition assay using capillary tubes.
- Variations in medium pH (6.4-8.2).
- Addition of varying concentrations of calf foetal serum and autologous serum.
- Manipulation of calcium ion (Ca++) concentration and use of calcium ionophore A 23187.
Main Results:
- pH variations between 6.4 and 8.2 did not significantly affect leukocyte migration.
- 5% calf foetal serum optimized conditions for leukocyte migration.
- Autologous serum contaminated the medium, complicating test interpretation due to cell dispersion.
- Calcium ion concentration changes had no significant effect unless combined with calcium ionophore A 23187 (0.5 microM).
Conclusions:
- Leukocyte migration assays are robust within a physiological pH range.
- Calf foetal serum is beneficial for leukocyte migration assays.
- Autologous serum should be avoided to ensure clear test interpretation.
- Calcium ions alone do not significantly modulate leukocyte migration; synergistic effects with ionophores are noted.
