title: GSM9221431: Lung immune ane epithelial cells from RA_2 patient; Homo sapiens; RNA-Seq
library_construction_protocol: Frozen lung tissues in cryovials were thawed at 37℃ for 2 minutes in a water bath, washed in Hank's balanced salt solution (HBSS), and then centrifuged for 5 minutes at 500g and 4℃. After that, lung tissues were put in 10cm petri dishes and finely minced. Subsequently, the minced tissues were incubated in HBSS containing 1X Liberase TM (0.25 U/mL) and DNaseI (0.5 mg/ml), and agitated at 360 rpm at 37℃ for 30 minutes. For enriching epithelial cells and immune cells, the dissociated single-cell suspensions were stained with antibodies for EPCAM, CD45, and CD31 and sorted using fluorescence-activated cell sorting (FACS) with a BD Influx cell sorter. The viability of the cells was determined by staining the cell preparation with DAPI 30 minutes prior to cell sorting. Library was performed according to the manufacter's protocol (10X Chromium GEM-X Single Cell 3' kit v4).