KEY RESOURCES TABLE

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Rabbit anti-TAGLN (SM22)	Abcam	Cat#ab14106 AB_443021
Goat anti-SOX9	R&D Systems	Cat#AF3075 AB_2194160
Rabbit anti-SOX9	Millipore	Cat#AB5535 AB_2239761
Rabbit anti-SOX9	Santa Cruz	Cat#sc-20095 AB_661282
Mouse anti-ECAD	BD Biosciences	Cat#610181 AB_397580
Goat anti-SOX2	R&D Systems	Cat#AF2018 AB_355110
Rabbit anti-SOX2	Seven Hills Bioreagents	Cat#WRAB-1236 AB_2715498
Rabbit anti-Non-phospho (Active) β-Catenin	Cell Signaling Technology	Cat#8814 AB_11127203
Rabbit anti-TTF1	Abcam	Cat#ab76013 AB_1310784
Goat anti-TP63	R&D Systems	Cat#BAF1916 AB_2207173
Rabbit anti-CHGA	Abcam	Cat#ab15160 AB_301704
Mouse anti-RAGE	Abcam	Cat#ab54741z AB_2242462
Mouse anti-ABCA3	Seven Hills Bioreagents	Cat#WMAB-17G524 AB_577285
Mouse anti-FOXJ1	Seven Hills Bioreagents	Cat#WMAB-319 AB_451719
Rabbit anti-FLAG	Sigma	Cat#F2555 AB_796202
Rabbit anti-KI67	Thermo Fisher	Cat#RM-9106-S1 AB_149792
Rabbit anti-CCAS3	Cell Signaling	Cat#9664 AB_2070042
Rabbit anti-pro-SP-C	Seven Hills Bioreagents	Cat#WRAB-9337 AB_2335890
Mouse anti-SP-B	Seven Hills Bioreagents	Cat#WMAB-1B9 AB_451723
Rabbit anti-SCGB3A2	Abcam	Cat# ab181853
Mouse anti-HTII-280	Terrace Biotech	Cat#TB-27AHT2-280 AB_2832931
Mouse anti-alpha smooth muscle actin	Sigma	Cat#C6198 AB_476856
Mouse anti-MUC5AC	Abcam	Cat#ab79082 AB_1603327
LIFR alpha PE	R&D Systems	Cat#FAB249P AB_2136109
IgG1 PE isotype control	R&D Systems	Cat#IC002P AB_357242
CD326 (EPCAM) FITC	Miltenyi	Cat#130-113-825 AB_2726341
REA control IgG1 FITC	Miltenyi	Cat#130-113-437 AB_2733689
CD31 APC	Miltenyi	Cat#130-117-314 AB_2727917
REA control IgG1 APC	Miltenyi	Cat#130-113-438 AB_2733893
Mouse anti-CD31 647	BD Biosciences	Cat#558094 AB_397020
Goat anti-mouse IgG2a FC APC	Thermo Fisher	Cat#31983
Bacterial and virus strains
LGR5 ECD adenovirus	(Yan et al., 2017)	N/A
Control adenovirus	(Yan et al., 2017)	N/A
pLKO.1 all-species scrambled non-target shRNA lentivirus	University of Michigan Vector Core; shRNA originally from Sigma Aldrich	Cat#SHC016
pLKO.1 human LGR5 shRNA lentivirus	University of Michigan Vector Core; shRNA originally from Sigma Aldrich	Clone: TRCN0000011586
Biological samples
Human fetal lung (8.5 – 19 weeks post-conception)	University of Washington Laboratory of Developmental Biology	N/A
Chemicals, peptides, and recombinant proteins
Collagen Type I	Thermo Fisher	Cat#A10483-01
Human fibronectin	Corning	CaT#356008
B-27 Supplement	Thermo Fisher	Cat#17504044
N-2 Supplement	Thermo Fisher	Cat#17502048
1-Thioglycerol	Sigma	Cat#M1753
Red Blood Cell Lysis Buffer	Sigma	Cat#11814389001
Normal donkey serum	Sigma	Cat#D9663
Tween 20	Thermo Fisher	Cat#BP337
Trisodium citrate	Sigma	Cat#S1804
CHIR99021	APExBIO	Cat#A3011
Recombinant human FGF7	R&D Systems	Cat#251-KG
All trans retinoic acid	Sigma	Cat#R2625
L-Ascorbic acid	Sigma	Cat#A4544
Y-27632	APExBIO	Cat#A3008
Recombinant human RSPO2	R&D Systems	Cat#3266-RS
Afamin/WNT3a conditioned media	MBL International Corporation	Cat#J2-001
Dispase (for bud tip organoid establishment)	Corning	Cat#354235
Dispase (for FACS)	Thermo Fisher	Cat#17105041
Collagenase Type II	Thermo Fisher	Cat#17101015
TrypLE Express Enzyme	Thermo Fisher	Cat#12605010
Polybrene Transfection Reagent	Sigma	Cat#TR-1003-G
Puromycin dihydrochloride	Sigma	Cat#P9620
Critical commercial assays
Neural Tissue Dissociation Kit	Miltenyi	Cat#130-092-628
RNAscope Multiplex Fluorescent Reagent Kit v2	ACD Bio	Cat#323100
MagMAX-96 Total RNA Isolation Kit	Thermo Fisher	Cat#AM1830
PicoPure RNA Isolation Kit	Thermo Fisher	Cat#AM1830
SuperScript VILO cDNA Kit	Thermo Fisher	Cat#11754250
QuantiTect SYBR Green PCR Kit	Qiagen	Cat# 204145
Deposited data
Raw scRNA-seq data from in vivo human fetal lung	(Miller et al., 2020)	ArrayExpress: E-MTAB-8221
Raw scRNA-seq data from lung explants	This manuscript	ArrayExpress: E-MTAB-10662
Experimental models: cell lines
Human fetal lung explants originated from fresh tissue sourced from the University of Washington Laboratory for Developmental Biology	N/A	N/A
Bud tip organoids were derived from fresh tissue sourced from the University of Washington Laboratory for Developmental Biology	N/A	N/A
Human fetal lung mesenchymal cells were isolated from fresh tissue sourced from the University of Washington Laboratory for Developmental Biology	N/A	N/A
Experimental models: organisms/strains
Oligonucleotides
For primer sequences, see Table S2	IDT	N/A
Recombinant DNA
Software and algorithms
CellProfiler	(Lamprecht, Sabatini and Carpenter, 2007)(Jones et al., 2008; Erben et al., 2018)	https://cellprofiler.org
Prism	GraphPad	https://www.graphpad.com/scientific-software/prism/
Sony Sorter MA900 associated software	Cytosens	https://www.cytosens.com/2020/03/25/how-you-can-easily-sort-fluorescent-protein-expressing-cells/
Scanpy	(Wolf, Angerer and Theis, 2018)	https://github.com/theislab/scanpy
CellRanger	10x Genomics	https://support.10xgenomics.com/single-cell-gene-expression/software/pipelines/latest/what-is-cell-ranger
Python	Python	Python.org
Methods and code for scRNA-seq analysis	GitHub	https://github.com/jason-spence-lab/Hein_2021
Other
Nucleopore Track-Etched Membranes	Sigma	Cat#WHA110414
Matrigel	Corning	Cat#354234
RNAse Away	Thermo Fisher	Cat#700511
Histo-Clear II	National Diagnostics	Cat#HS-202
ProLong Gold	Thermo Fisher	Hs-LGR6
TSA Plus Cyanine 5	Akoya Biosciences	Cat#NEL705001KT
TSA Plus Cyanine 3	Akoya Biosciences	Cat#NEL744001KT
Hs-RSPO2-02	ACD Bio	Cat#423991
Hs-LGR5	ACD Bio	Cat#311021
Hs-AXIN2	ACD Bio	Cat#400241
Hs-PDGFRA-C2	ACD Bio	Cat#604481-C2
Hs-FGFR4-no-XMm-C2	ACD Bio	Cat#443431-C2
Hs-WNT2	ACD Bio	Cat#584071
Hs-EGR1-C2	ACD Bio	Cat#457671-C2
Hs-RSPO1	ACD Bio	Cat#446921
Hs-RSPO3-01	ACD Bio	Cat#429851
Hs-RSPO4	ACD Bio	Cat#510201
Hs-LGR4	ACD Bio	Cat#460551
Hs-LGR6	ACD Bio	Cat#410461
Hs-SCGB3A2	ACD Bio	Cat#549951
Hs-SFTPB	ACD Bio	Cat#544251
Hs-AGTR2	ACD Bio	Cat#459141
Hs-BMP4-C2	ACD Bio	Cat#45301-C2
Hs-SERPINF1-C3	ACD Bio	Cat#564391-C3