FACS - based isolation of human eosinophils allows purification of high quality RNA.
Fiche publication
Date publication
septembre 2018
Journal
Journal of immunological methods
Auteurs
Membres identifiés du Cancéropôle Est :
Pr BAHRAM Siamak, Pr GENY Bernard
Tous les auteurs :
Wacht G, Poirot A, Charles AL, Radosavljevic M, Uring-Lambert B, de Blay F, Geny B, Bahram S, Barnig C
Lien Pubmed
Résumé
Here we extensively describe a FACS-based protocol for isolating intact non-stained human eosinophils from peripheral blood; a stop forward from our recently published initial study. This method of purification could be accomplished in <3 h with only small volumes of whole blood necessary, even in healthy subjects generally exhibiting low levels of circulating eosinophils. Eosinophil activation during the isolation steps appeared to be minimal and this purification procedure yielded high quality RNA. Moreover, these FACS-isolated eosinophils had prolonged viability in culture and were suitable for further activation assays.
Mots clés
Eosinophils, Fluorescence-activated cell sorting (FACS), RNA, Superoxide release
Référence
J. Immunol. Methods. 2018 Sep 11;: