GSM4838954: mTEC2_thymic epithelial cells_scRNA-seq; Homo sapiens; RNA-Seq
Thymic tissue fragments were dissociated to single cells by enzymatic treatment (0.4 mg/mL Collagenase D (Roche), 0.6 mg/mL Dispase II (Roche), 40 μg/mL DNAse I (Roche)) for about 30-45 minutes at 37ºC. Cells were pelleted and re-suspended for cell counting. single cell suspension was depleted of CD45 and CD235ab (red blood) expressing cells by immunomagnetic separation. The enriched fraction was stained and sorted using FACSAria III machine for cortical (EpCAMlowCD205+) and medullary  (EpCAMhighCD205-) thymic epithelial cells (TEC) RNA libraries were prepared for sequencing using Chromium Single Cell 3' v2 protocol (PN-120233, 10X Genomics); cDNA amplification involved 12 PCR cycles