GSM8707303: untreated-F; Mus musculus; RNA-Seq
Freshly harvested lung tissue from C57BL/6J mice was minced to a slurry and suspended in complete media (Dulbecco's modified Eagle's medium [DMEM], 10% FBS, 1% Gibco™ Antibiotic-Antimycotic) containing collagenase (1 mg/ml) and DNase I (0.25 ml or 250 KU units). The suspension was incubated on a rocker for 30 min at 37°C followed by centrifugation at 500 x g for 10 min. The supernatant was decanted, and each pellet was briefly resuspended in sterile water for RBC lysis, neutralized by adding 10 ml of complete media. Cells were centrifuged and pellets resuspended in 5 ml complete media and passed through a 100 μm nylon screen to obtain a single-cell suspension. Cells were centrifuged and resuspended in PBS. FACS was used to isolate live cells (absence of DAPI staining). Library was generated  according to the manufacturer's protocol CG000315 Rev C for the Chromium Next GEM Single Cell 3' Kit v3.1 (10X Genomics, Pleasanton, CA, USA).