GSM8477281: Ctrl, replicate 3; Mus musculus; RNA-Seq
Lung tissues were harvested from mice and incubated with digestion media (RPMI, 2mg/ml collagenase A, and 0.1mg/ml DNaseI) at 37°C for 20 minutes on a rocker. The resulted single cell suspension was filtered through a 70um filter, then subsequently incubated with ACK lysis buffer for 3 minutes. Cells were stained with viability dye and were resuspend in FACS staining buffer (PBS, 2% FBS, 0.1% sodium azide). Cells were then stained with conjugated anti-CD45 and antiEpcam antibody for 30 minutes at 4°C. Cells then were sorted to live CD45+/Epcam- populations. For each sample, 10,000 cells were collected for scRNA-seq. Libraries were generated with Chromium Single Cell 3' Reagent Kits.