Isolation of human eosinophils: microbead method has no impact on IL-5 sustained viability

人嗜酸性粒细胞分离:微珠法对IL-5持续活力无影响

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作者:Caroline M Percopo, Kimberly D Dyer, Kristin E Killoran, Helene F Rosenberg

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.

Methods

We isolated eosinophils from normal donor granulocyte packs using two methods, and evaluated purity, viability and annexin-V/propidium-iodide staining.

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.

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.

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