Eosinophils scRNA-seq from healthy and asthmatic donors
Eosinophils scRNA-seq from healthy and asthmatic donors
Eosinophils were purified from the peripheral blood of healthy controls and asthmatic donors. For eosinophil purification, CD2, CD3, CD14, CD16, CD19, CD20, CD36, CD56, CD123, and glycophorin A positive cells were discarded by a magnetic bead separation technique, as described in the manufacturer's procedure (EasySep; StemCell Technologies, Vancouver, Canada). Eosinophils whole transcriptome analyses were performed using the BD Rhapsody™ Single Cell Analysis System (BD Biosciences). Purified eosinophils (120000 cells) were centrifuged and resuspended in 650 μL of BD sample buffer supplemented with 1:1000 SUPERase-In (20 U/μL; AM2694 ThermoFisher) and NxGen Rnase Inhibitors (40 U/μL; 30281-2 Lucigen), as previously described. For each sample, 20000-40000 cells were loaded into a different Rhapsody Single-Cell Analysis System cartridge. Cell capture and cDNA synthesis were performed according to manufacturer instructions.