GSM5850285: RCDII-2; Homo sapiens; RNA-Seq
Intraepithelial lymphocytes (IELs) were isolated from duodenal biopsies by treatment with HBSS/1 mM EDTA (made in-house; J.T. Baker) rolling for 2x 45 min at 37°C. Cells were collected in 20% FCS/RPMI and kept at 4°C. Subsequently, single-cell suspensions were obtained from the lamina propria (LP) by  1.5 hours enzymatic treatment with 10 units/mL collagenase IV (Worthington) and 200 µg/ml DNase I grade II (Roche). For obtainment of single cells, LP cells and IELs were resuspended and passed through a 70 µm cell strainer. Cells were washed with 0.5% FCS/DPBS (Dulbecco's Phosphate Buffered Saline) (Gibco, Thermo Fisher) by centrifuging at 1650 rpm for 10 min. Hereafter, cells were treated with red blood cell lysis buffer (LUMC pharmacy) and removal of dead cells was performed using microbeads (Miltenyi Biotec). Lastly, live CD45+ cells were positively selected using MACS beads (Miltenyi Biotec). Chromium 3' Single cell platform (10X Genomics)