GSM7927623: Mouse_control_lung_batch_1; Mus musculus; RNA-Seq
The lungs were dissociated using digestion buffer in C-tube (Miltenyi Biotec). After 30 min at 37 °C, the samples were treated with final GentleMACS dissociation. Cell suspensions were passed through a 70-μm cell strainer. Lung mononuclear cells were separated using Percoll solution (GE Healthcare). CD4+ or CD45+ cells were stained and freshly isolated using a FACSAria III cell sorter (BD Biosciences). For human samples, Freshly obtained nasal polyps were immediately minced and incubated in RPMI 1640 medium containing 1 mg/ml collagenase, 0.5 mg/ml hyaluronidase, and 0.2 mg/ml DNase I (Sigma-Aldrich). After incubation, nasal polyp mononuclear cells were purified by density gradient centrifugation using Percoll solution (GE Healthcare). PBMCs were isolated from peripheral blood using Ficoll-Paque Plus solution (GE Healthcare). CD4+ T cells were stained and sorted using a FACSAria III cell sorter (BD). Libraries were prepared using Chromium Single Cell 3' Reagent Kits v3.1 according to the manufacturer's protocol (10X Genomics). The generated scRNA-seq libraries were sequenced using a 128 cycle (paired-end reads) with a NovaSeq 6000 (Illumina).