scRNA_E130ESO_Epi
Esophageal tubes were cut open using fine scissors, and gently digested in 6 cm dishes with 25 U mL Dispase (Corning) + 100 μg mL DNase I (Millipore Sigma) at room temperature for 20 to 40 min. Digestion was stopped by transferring esophagus to cold HBSS. Epithelium was physically peeled off from surrounding stromal tissues with fine forceps, and collected in a 15 ml Falcon tube. The epithelium was further dissociated into single cells in TrypLE Express (Thermo Fisher Scientific), first triturated gently using P1000 pipette and then shaken in a Thermomixer at 37 ​​°C at 1000 rpm for 10 to 20 min. Dissociation was stopped by addition of the FACS buffer (PBS + 1% BSA + 2 mM EDTA + 1X Anti-Anti + 25 mM HEPES pH 7.0) supplemented with 10% FBS. Cells were transferred to 1.5 ml Eppendorf tubes and pelleted down at 300 g at 4 °C for 5 min. Then cells were resuspended in cold 0.04% BSA in PBS for cell number counting. Cell alive rate was determined using Trypan Blue. Only samples with greater than 80% cell alive rate were further processed. Cells were then subjected to 10X Chromium scRNA-Seq based on the manual.