Lineage Tracing of PSC-derived iAEC2 Differentiation Using Lentiviral Barcoding

Lineage tracing of individual cells during directed differentiation human iPSC into alveolospheres was performed using a lentiviral barcode labeling system (Weinreb et al., 2020) as described in Hurley et al., 2020. Human BU3 NGST iPSCs were differentiated in vitro, sorted on day 15 for NKX2-1GFP positive cells, which were grown in CK+DCI until D17 (as described in Hurley et al., 2020). At this point cells were infected with a lentiviral barcode to demarcate individual cells. Cells were then replated and grown out for an additional 10 days before being analyzed by single cell sequencing.