Single-cell analysis of human lung epithelia reveals concomitant expression of the SARS-CoV-2 receptor ACE2 with multiple virus receptors and scavengers in alveolar type II cells

Guangchun Han; Ansam Sinjab; Warapen Treekitkarnmongkol; Patrick Brennan; Kieko Hara; Kyle Chang; Elena Bogatenkova; Beatriz Sanchez-Espiridion; Carmen Behrens; Boning Gao; Luc Girard; Jianjun Zhang; Boris Sepesi; Tina Cascone; Lauren Byers; Don L. Gibbons; Jichao Chen; Seyed Javad Moghaddam; Edwin J. Ostrin; Junya Fujimoto; Jerry Shay; John V. Heymach; John D. Minna; Steven Dubinett; Paul A. Scheet; Ignacio I. Wistuba; Edward Hill; Shannon Telesco; Christopher Stevenson; Avrum E. Spira; Linghua Wang; Humam Kadara

Samples were then filtered through 70 μm strainers (Miltenyi biotech, 130-098-462) and washed with ice-cold HBSS. Filtrates were then centrifuged and re-suspended in ice-cold ACK lysis buffer (A1049201, Thermo Fisher Scientific) for red blood cell (RBC) lysis. Following RBC lysis, samples were centrifuged and re-suspended in ice-cold FBS, filtered (using 40 μm FlowMi tip filters; H13680-6 FBS) for 30 minutes. EPCAM-stained cells were then washed, filtered using 40 μm tip filters, stained with SYTOX Blue and processed on a FACS Aria II instrument. Doublets and dead cells were eliminated, and viable (SYTOX-negative) EPCAM-positive (EPCAM+, epithelial) singlets were collected in PBS containing 2% FBS. Cells were washed again to eliminate ambient RNA, and a sample was taken for counting by Trypan Blue exclusion (T8154, Sigma Aldrich) before loading on 10X Chromium microfluidic chips.