Single-site, five-year experience with human eosinophil isolation by density gradient centrifugation and CD16...

Area of Science:

• Hematology
• Immunology
• Cell Biology

Background:

• Reliability of human eosinophil purification methods is not well-documented.
• Eosinophils play a crucial role in allergic inflammation and immune responses.
• Accurate isolation of eosinophils is essential for research and clinical applications.

Purpose of the Study:

• To evaluate the reliability and reproducibility of immunomagnetic separation for human eosinophil isolation.
• To assess the yield, purity, and viability of isolated eosinophils.
• To compare eosinophil isolation outcomes between allergic and non-allergic donors.

Main Methods:

• Retrospective review of 350 consecutive eosinophil isolations using immunomagnetic separation.
• Analysis of absolute eosinophil count (AEC) from complete blood counts.
• Assessment of eosinophil yield, purity, and viability post-isolation.
• Reproducibility analysis in donors with multiple blood draws.

Main Results:

• Immunomagnetic separation yielded an average of 3.1 million eosinophils per 20 mL of blood with >98% purity.
• Eosinophil recovery averaged 87% ± 29%, strongly correlating with AEC.
• Isolated eosinophil viability was consistently >95% and did not differ between allergic and non-allergic donors.
• The method demonstrated reproducibility across multiple draws from the same donors.

Conclusions:

• Immunomagnetic separation is a reliable and reproducible technique for isolating human eosinophils from whole blood.
• The method consistently achieves high purity (>98%) and viability (>95%) of eosinophils.
• This technique provides a dependable approach for obtaining sufficient eosinophils for research purposes.