Homo sapiens
Protocols: Distal regions of 12 week fetal lungs were cut into ~2mm sections and incubated with dispase, 100% FBS and then 10% FBS as described above, and moved to a sterile petri dish. Mesenchyme was removed by repeated pipetting of distal lung pieces after dispase treatment. Buds were washed with DMEM until mesenchymal cells were no longer visible. Buds were then moved to a 1.5 mL eppendorf tube containing 200 µL of Matrigel, mixed using a p200 pipette, and plated in ~20 µL droplets in a 24 well tissue culture plate. Plates were placed at 37 degrees Celsius with 5% CO2 for 20 minutes while droplets solidified. 500uL of media was added to each well containing a droplet. Media was changed every 2-4 days. RNA was isolated using the mirVana RNA isolation kit, following the “Total RNA” isolation protocol (Thermo-Fisher Scientific, Waltham MA). RNA sequencing library preparation and sequencing was carried out by the University of Michigan DNA Sequencing Core and Genomic Analysis Services (https://seqcore.brcf.med.umich.edu/).