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A rapid simple method of basophil purification by density centrifugation on Percoll
Journal of Immunology (Baltimore, Md. : 1950)
|November 1, 1982
Summary
Researchers developed rapid methods to purify basophils, a type of white blood cell, from human blood using Percoll density gradients. This technique isolates viable basophils for further study, though yields are currently limited.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Basophils are crucial white blood cells involved in immune responses.
- Efficient isolation of pure basophils is essential for studying their function and role in diseases.
- Existing methods for basophil purification can be time-consuming or yield low purity.
Purpose of the Study:
- To establish simple, rapid, and effective methods for purifying basophils from human peripheral blood.
- To determine optimal Percoll densities for selective basophil concentration.
- To assess the viability and functional integrity of isolated basophils.
Main Methods:
- Isopyknic centrifugation of anticoagulated human peripheral blood over Percoll density gradients.
- Collection of basophil-rich cell fractions based on density distribution.
- Morphological and functional assessments (histamine release) of purified cells.
- Optional second centrifugation step for enhanced purity.
Main Results:
- Basophils were primarily found at densities between 1.072 and 1.078 g/ml, peaking at 1.074-1.076 g/ml.
- A single Percoll gradient step achieved 24.4% +/- 4% basophil purity.
- Isolated basophils were viable, morphologically normal, and responsive to stimuli.
- Two-step purification enhanced purity but resulted in low basophil recovery (<= 20%).
Conclusions:
- Simple and rapid Percoll density gradient centrifugation effectively isolates viable human basophils.
- The method provides a valuable tool for basophil research, despite current yield limitations.
- Further optimization is needed to improve the recovery rate of purified basophils.