Isolation of human eosinophils: microbead method has no impact on IL-5 sustained viability
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Updated: Jun 20, 2026
Isolation of Human Neutrophils from Whole Blood and Buffy Coats
Published on: September 17, 2021
Background/Purpose:
In a recent issue of Experimental Dermatology (18, 2009, 654), Schefzyk et al. concluded that multi-antibody eosinophil isolation (Miltenyi) should be abandoned, as differential purity was minimal, and eosinophils underwent accelerated apoptosis when compared with those isolated with traditional anti-CD16 microbeads. Our intent was to investigate the universality of these findings.
Methods:
We isolated eosinophils from normal donor granulocyte packs using two methods, and evaluated purity, viability and annexin-V/propidium-iodide staining.
Results:
Purity was substantially greater when multi-antibody isolation was used for eosinophil isolation from granulocyte packs (98% vs 69%). No differential survival was detected when eosinophils were maintained in culture with or without interleukin-5.
Conclusions:
Multi-antibody eosinophil isolation represents a substantial advantage over anti-CD-16 microbeads when isolating large numbers of eosinophils from concentrated leucocyte preparations. No differential survival was observed. While appropriate consideration of methods is always crucial, multi-antibody eosinophil isolation should not be abandoned completely.
Insights
Multi-antibody eosinophil isolation yields higher purity and comparable viability to anti-CD16 microbeads. This method offers a significant advantage for isolating large numbers of eosinophils, suggesting it should not be abandoned.
Area of Science:
- Immunology
- Cell Biology
Background:
- Previous research suggested abandoning multi-antibody eosinophil isolation due to low purity and accelerated apoptosis.
- These findings contrasted with traditional anti-CD16 microbead isolation methods.
Purpose of the Study:
- To investigate the universality of previous findings on multi-antibody eosinophil isolation.
- To compare the purity and viability of eosinophils isolated using multi-antibody versus anti-CD16 microbead methods.
Main Methods:
- Eosinophils were isolated from normal donor granulocyte packs using two distinct methods.
- Purity and viability were assessed, including annexin-V/propidium-iodide staining.
Main Results:
- Multi-antibody isolation resulted in substantially greater eosinophil purity (98%) compared to anti-CD16 microbeads (69%).
- No significant difference in eosinophil survival was observed between the two isolation methods, even with interleukin-5 stimulation.
Conclusions:
- Multi-antibody eosinophil isolation offers a significant advantage for isolating large quantities of eosinophils from concentrated leukocyte preparations.
- The findings do not support completely abandoning multi-antibody eosinophil isolation, highlighting its utility.
- Methodological considerations remain important in cell isolation techniques.
