Purification of human blood basophils using negative selection by flow cytometry
Y Tanimoto1, K Takahashi, M Takata
1Second Department of Internal Medicine, Okayama University Medical School, Japan.
Summary
Researchers developed a method to purify basophils, a type of white blood cell, from blood. This technique preserves basophil function, making it valuable for studying allergic reactions like asthma.
Area of Science:
- Immunology
- Cell Biology
Background:
- Basophils play a crucial role in hypersensitivity reactions.
- Accurate in vitro studies require highly purified and functional basophils.
Purpose of the Study:
- To establish a reliable method for purifying human basophils.
- To assess if the purification process affects basophil function.
Main Methods:
- Purification of basophils from peripheral blood using Percoll discontinuous gradients and negative selection via flow cytometry.
- Assessment of basophil purity, yield, and viability.
- Evaluation of histamine content, spontaneous and triggered histamine release, and chemotactic response to complement C5a.
Main Results:
- Achieved high basophil purity (mean 84.7%) and viability (>90%).
- Purification method did not alter intracellular histamine levels or histamine release responses.
- Purified basophils demonstrated dose-dependent chemotaxis to complement C5a.
Conclusions:
- The described purification procedure yields highly pure, functional human basophils.
- This method is suitable for in vitro research on basophil roles in allergic diseases like asthma.
