GSM7776447: Donor 4; Homo sapiens; RNA-Seq
Thymocytes were thawed at room temperature in RPMI 1640 containing FCS 30%, Penicillin-Streptomycin 1%, DNase I (10 ug/ml, Sigma), and heparin 20 U/ml. After washing, FACS surface staining was performed on ice, after FC blockage, using the following antibodies: Vα24 (FITC, Beckman Coulter), Vβ11 (APC, Beckman Coulter), CD3 (BV605, Biolegend), CD14 (APC Cy7, Biolegend), CD19 (APC Cy7, Biolegend), LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit (Invitrogen). CD3+, Vα24+, Vβ11+, CD14-, CD19- live cells were FACS-sorted on a FACS Aria-III (BD Biosciences). Single-cell libraries were generated from FACS-sorted human thymic iNKT cells using the Chromium Controller Single-Cell Instrument and Chromium X instrument and Chromium Single 3′ Library & Gel Bead Kit v3 (10x Genomics).