Human embryonic lung epithelial tips are multipotent progenitors that can be expanded in vitro as long-term self-renewing organoids

  1. Marko Z Nikolić
  2. Oriol Caritg
  3. Quitz Jeng
  4. Jo-Anne Johnson
  5. Dawei Sun
  6. Kate J Howell
  7. Jane L Brady
  8. Usua Laresgoiti
  9. George Allen
  10. Richard Butler
  11. Matthias Zilbauer
  12. Adam Giangreco
  13. Emma L Rawlins  Is a corresponding author
  1. Wellcome Trust/CRUK Gurdon Institute, University of Cambridge, United Kingdom
  2. University of Cambridge, United Kingdom
  3. European Molecular Biology Laboratory, European Bioinformatics Institute, Wellcome Genome Campus, United Kingdom
  4. University of Cambridge and Addenbrookes Hospital, United Kingdom
  5. University College London, United Kingdom
  6. Wellcome Trust/MRC Stem Cell Institute, United Kingdom
6 figures, 2 videos, 9 tables and 2 additional files

Figures

Figure 1 with 5 supplements
Evolution of alveolar and bronchiolar marker gene expression during human embryonic lung development.

Sections of human embryonic and adult lungs. (A, B) 11 and 20 pcw. Green: HTII-280; red: SOX9 (tips); white: ECAD (epithelial cells). Arrow = HTII-280 positive terminal airway. (C) Adult. Green: …

https://doi.org/10.7554/eLife.26575.003
Figure 1—figure supplement 1
Representative morphology of human embryonic lung samples.

Wholemount images of human embryonic lungs. (A) CS15 (~5 pcw). (B) 6 pcw. (C) 8 pcw. (D) 10 pcw. (E) 16 pcw. Bars = 2.5 mm (A, B); 5 mm (C, D); 3 cm (E).

https://doi.org/10.7554/eLife.26575.004
Figure 1—figure supplement 2
The evolution of alveolar marker gene expression in the human embryonic lung.

Sections of human embryonic and adult lungs. (A) 14 and 17 pcw. Green: HTII; red: SOX9 (tips); white: ECAD (epithelial cells). (B) 14 and 17 pcw. Green: HTII-280; red: HOPX; white: SOX2 (bronchiolar …

https://doi.org/10.7554/eLife.26575.005
Figure 1—figure supplement 3
Epithelial PDPN expression is not specific to AT1 cells during human lung development.

Sections of human and adult embryonic lungs. (A, A’) nine pcw. Green: NKX2-1; red: PDPN. Arrowheads = low levels of PDPN in the developing airways. Arrows = PDPN in lymphatic endothelial cells. (B) …

https://doi.org/10.7554/eLife.26575.006
Figure 1—figure supplement 4
Epithelial AQP5 expression is not specific to AT1 cells during human lung development.

Sections of human and adult embryonic lungs. (A–D) 11, 14, 15 and 20 pcw. Green: AQP5; red: ECAD (epithelial cells). Patches of diffuse, apical AQP5 staining can be observed from 11 pcw. Arrowheads …

https://doi.org/10.7554/eLife.26575.007
Figure 1—figure supplement 5
NKX2-1 and FOXA2 are expressed in all human lung epithelial cells up to 20 pcw.

Sections of human embryonic lungs. (A–D). 15, 17, 20 and 21 pcw. Green: NKX2-1; red: FOXA2; white: HTII-280. Arrowheads in D’ = NKX2-1- cells apparently lining the developing alveolar sacs. E-G. 20 …

https://doi.org/10.7554/eLife.26575.008
Figure 2 with 5 supplements
The tip and stalk epithelial cell populations are clearly demarcated in branching human, pseudoglandular stage, lungs.

(A–E) Sections of human embryonic lungs. (A) 11 pcw. Green: SOX9 (tip); red: SOX2 (stalk); white: α-SMA (smooth muscle). (B) 8 pcw. Green: SOX9 (tip); red: SOX2 (stalk); white: KI67 (proliferating …

https://doi.org/10.7554/eLife.26575.009
Figure 2—figure supplement 1
Pseudoglandular stage human lung tips co-express SOX9 and SOX2.

(A) E13.5 mouse lung staining illustrating absence of SOX2 in SOX9+ tip cells. Green: SOX9; red: SOX2; white: ECAD. (B) Whole-mount staining of a 5 pcw human embryonic lung showing the primary …

https://doi.org/10.7554/eLife.26575.010
Figure 2—figure supplement 2
Human lung tips down-regulate SOX2 during the canalicular stage.

(A–G) Sections of human embryonic lungs at 11, 14, 17 and 20 pcw stained for green: SOX9; red: SOX2. Slides were stained simultaneously and imaged using the same microscope settings such that …

https://doi.org/10.7554/eLife.26575.011
Figure 2—figure supplement 3
Transcriptional differences and similarities between human pseudoglandular stage tip and stalk populations.

(A) Chart to show the percentage of the expressed genes in the major gene ontology classes represented in the data. (Note: tip and stalk data are reproduced from Figure 2I). (B–E) Human embryonic …

https://doi.org/10.7554/eLife.26575.012
Figure 2—figure supplement 4
Specific transcription factors are enriched at the protein level in human distal epithelial lung tips.

(A–D) Sections of 8 pcw human embryonic lung. (A) Green: ETV5; red: SOX2; white: ECAD (epithelial cells). (B) Green: HMGA2; red: SOX2; white: ECAD (epithelial cells). (C) Green: HNF1B; red: SOX2; …

https://doi.org/10.7554/eLife.26575.013
Figure 2—figure supplement 5
Tip gene expression is highly conserved between mouse and human.

(A) Pie chart showing over-lap of orthologous human-mouse genes called as present in 6–7 pcw human tips and E11.5 mouse tips. (B) Scatter plot to estimate the relative levels of expression of …

https://doi.org/10.7554/eLife.26575.014
Figure 3 with 5 supplements
Long-term, self-renewing organoid culture of human lung epithelial tip cells with a initial colony forming efficiency of 100%.

(A) Frames from Video 1 showing bright field images of a single microdissected tip taken every 24 hr for 12 days. Representative bright field images of tip organoid cultures from P0, P6 and P15. A …

https://doi.org/10.7554/eLife.26575.015
Figure 3—figure supplement 1
Organoids passaged in self-renewing medium retain tip-specific transcription factor proteins.

(A–F) Cryosections of self-renewing tip organoids between passage 5 and 7. (A) Green: SOX9; red: SOX2; white: ECAD (epithelial cells). (B) Green: HMGA2; red: SOX2; white: ECAD (epithelial cells). (C)…

https://doi.org/10.7554/eLife.26575.016
Figure 3—figure supplement 2
Organoids passaged in self-renewing medium retain a normal karyotype.

(A–G) Karypotype of 7 organoid lines between passage 4 and 8. 7/7 organoid lines tested retained a normal karypotype.

https://doi.org/10.7554/eLife.26575.017
Figure 3—figure supplement 3
SOX2-, SOX9+ canalicular stage human embryonic tips can be grown as SOX2+, SOX9+self-renewing organoids.

(A–D) Bright field images of 19 pcw tips growing as self-renewing organoids. (E) 19 pcw tip organoids co-express SOX2 and SOX9. Green: SOX9; red: SOX2; white: ECAD. Blue: DAPI. Scale bars = 2 mm (A–D

https://doi.org/10.7554/eLife.26575.018
Figure 3—figure supplement 4
Box plots of selected tip and stalk specific genes showing transcript levels in fresh tissue and cultured organoids.

Normalised RNAseq data was used to generate box plots of selected genes.

https://doi.org/10.7554/eLife.26575.019
Figure 3—figure supplement 5
E12.5 mouse tips do not long-term self-renew in the growth medium developed for human tips.

(A, C) Bright-field images of mouse E12.5 tip epithelium growing as organoids in self-renewal medium +/- SB431542. (Factors adjusted to be mouse-specific). (B, D) Confocal images of mouse organoids. …

https://doi.org/10.7554/eLife.26575.020
All factors added to the medium are required for culture establishment.

(A) Whole-mount staining for DAPI and ECAD was performed to estimate the fraction of mesenchyme in a microdissected tip (dotted area) and stalk (dashed box). (B) Quantitation of percentage of …

https://doi.org/10.7554/eLife.26575.023
Figure 4—source data 1

Individual data points for Figure 4B.

Percentage of cells scored as mesenchyme in freshly dissected tips and stalks. Expressed as percentage of non-ECAD+ cells from total DAPI+ cells. Quantitation done using the mesenchyme-macro.ijm for FIJI which is available as a supplemental file.

https://doi.org/10.7554/eLife.26575.024
Figure 4—source data 2

Individual data points for Figure 4F.

Percentage organoid forming efficiency and size increase from D1-11 of culture when fresh tips are plated in self-renewing medium +/- SB43125. Three independent experiments were performed on different lung samples.

https://doi.org/10.7554/eLife.26575.025
Figure 4—source data 3

Raw qRT-PCR data for Figure 4G.

Fold-change was normalised to one for self-renewing medium. Data for three individual organoid lines are shown.

https://doi.org/10.7554/eLife.26575.026
Figure 5 with 2 supplements
Disscoiated self-renewing organoids are competent to differentiate under the kidney capsule in the presence of embryonic mouse lung cells.

(A) Experimental schematic. Dissociated organoids were mixed with dissociated E13.5 mouse lungs (either MF1 outbred strain for grafts harvested at 3 and 7 weeks, or Rosa26R-mT-mG strain for grafts …

https://doi.org/10.7554/eLife.26575.027
Figure 5—figure supplement 1
Cells isolated from self-renewing organoids are competent to engraft into adult mouse lungs.

(A) Oropharyngeal administration of 1 mg/ml bleomycin at a dose of 1 μl per gram body weight is sufficient to injure adult male NSG mouse lungs. Representative haematoxylin and eosin stained lung …

https://doi.org/10.7554/eLife.26575.028
Figure 5—figure supplement 2
Mouse regions of chimeric human-mouse kidney capsule grafts differentiate efficiently.

The extent of differentiation of the mouse cells was indistinguishable at 3, 7 and 12 weeks post-grafting and a selection of representative images from the various times are shown. Regions of mouse …

https://doi.org/10.7554/eLife.26575.029
Figure 6 with 5 supplements
In vitro differentiation of self-renewing organoids towards bronchiolar and alveolar lineages.

(A) Experimental schematic for 3 weeks organoid differentiation in Pneumacult-ALI medium. (B) In low passage number organoids, mesenchyme expanded (arrows), basally-located TP63+ basal cells …

https://doi.org/10.7554/eLife.26575.030
Figure 6—figure supplement 1
Exposure of self-renewing organoids to PneumacultTM medium leads to efficient goblet cell differentiation with rare patches of KRT5+ basal cells.

(A) Experimental schematic. (B) Every organoid examined had many MUC5AC+ goblet cells and little or no SOX9. Green: MUC5AC (mucous); red: SOX9. (C) More rarely, some organoids contained patches of …

https://doi.org/10.7554/eLife.26575.031
Figure 6—figure supplement 2
Testing media conditions for ability to promote human organoid alveolar differentiation.

(A) Experimental schematic. (B) Alveolar differentiation media tested. DCI = dexamethasone, cAMP, IBMX. (C-D) Self-renewing controls co-express SOX2 and SOX9, whereas SOX2 is retained and SOX9 is …

https://doi.org/10.7554/eLife.26575.032
Figure 6—figure supplement 3
A combination of canalicular stage lung embryonic mesenchyme and alveolar differentiation medium together promote the most efficient organoid alveolar differentiation.

(A) Experimental schematic for 3 week alveolar differentiation of psedoglandular stage-derived organoids in the presence of freshly-isolated 19 pcw human mesenchyme. DCI = dexamethasone, cAMP, IBMX. …

https://doi.org/10.7554/eLife.26575.033
Figure 6—figure supplement 4
Expanded canalicular stage mesenchyme and alveolar medium can promote organoid alveolar differentiation.

(A) Experimental schematic for 3 week alveolar differentiation of psedoglandular and canalicular stage-derived organoids in the presence of expanded 19 or 20 pcw human mesenchyme. (B–D) …

https://doi.org/10.7554/eLife.26575.034
Figure 6—figure supplement 5
Expanded fibroblasts used for organoid co-cultures are a heterogeneous population expressing various lung embryonic fibroblast markers.

(A–D) 20 pcw lung sections stained for mesenchymal markers. (A) Green: PDGFRA; red: PDPN (alveolar epithelium and lymphatic endothelium); white: SMA. (B) Green: PDGFRB; red: PDPN (alveolar …

https://doi.org/10.7554/eLife.26575.035

Videos

Video 1
Human lung epithelial tip growing into an organoid over 11 days.

Imaged every 12 hr in bright-field on a Nikon Biostation.

https://doi.org/10.7554/eLife.26575.021
Video 2
Organoid structure is a single layer epithelium with a hollow lumen.

Confocal z-stack of P6 tip organoid. Nuclei (DAPI, blue) and epithelial cells (ECAD, white) illustrating typical organoid morphology.

https://doi.org/10.7554/eLife.26575.022

Tables

Table 1

Self-renewal (Human)

https://doi.org/10.7554/eLife.26575.036
ReagentCompanyCat noFinal concentration
Advanced DMEMThermoFisher Scientific, Invitrogen12634–010Base medium
Penicillin/StreptomycinThermoFisher Scientific, Invitrogen15140–122100 U/ml (Pen) 100 μg/ml (Strep)
HepesThermoFisher Scientific, Invitrogen15630–05610 mM
GlutamaxThermoFisher Scientific, Invitrogen35050–0382 mM
N2ThermoFisher Scientific, Invitrogen17502–0481:100
B27 (-Vit A)ThermoFisher Scientific, Invitrogen12587–0101:50
N-acetylcysteineSigma-AldrichA91651.25 mM
Matrigel (growth factor reduced; specific lots of matrigel with at least 8 mg/ml protein concentration were used)Corning356231undiluted
R-spondin1 conditioned mediumStem Cell Intitute, University of CambridgeFrom 293T-HA-Rspo1-Fc cell line made by Calvin Kuo, Stanford5% v/v
EGFPeprotech, UKAF-100–1550 ng/ml
NogginR and D Systems6057 NG-100100 ng/ml
FGF10R and D Systems345-FG-025100 ng/ml
FGF7Peprotech100–19100 ng/ml
CHIR 99021Stem Cell Institute, University of Cambridgen/a3 μM
SB 431542Tocris161410 μM
48 well plates (Greiner Cellstar)Sigma-AldrichM9437n/a
Table 2

Self-renewal (Mouse)

https://doi.org/10.7554/eLife.26575.037
ReagentCompanyCat noFinal concentration
Advanced DMEMThermoFisher Scientific, Invitrogen12634–010n/a
Penicillin/StreptomycinThermoFisher Scientific, Invitrogen15140–122100 U/ml (Pen)
100 μg/ml (Strep)
HepesThermoFisher Scientific, Invitrogen15630–05610 mM
GlutamaxThermoFisher Scientific, Invitrogen35050–0382 mM
N2ThermoFisher Scientific, Invitrogen17502–0481:100
B27 (-Vit A)ThermoFisher Scientific, Invitrogen12587–0101:50
N-acetylcysteineSigma-AldrichA91651.25 mM
MatrigelCorning356231Undiluted
R-spondin conditioned mediumStem Cell Institute, University of CambridgeFrom 293T-HA-Rspo1-Fc cell line made by Calvin Kuo, Stanford5% v/v
mEGFR and D Systems2028-EG-20050 ng/ml
NogginR and D Systems6057 NG-100100 ng/ml
FGF10R and D Systems345-FG-025100 ng/ml
mFGF7R and D Systems5028 KG_025100 ng/ml
CHIR 99021Stem Cell Institute, University of Cambridgen/a3 μM
SB 431542Tocris161410 μM
Table 3

Human foetal lung mesenchyme

https://doi.org/10.7554/eLife.26575.038
ReagentCompanyCat noFinal concentration
Dish Nunc T/C 92 mmThermoFisher Scientific10508921n/a
Type I collagenSigma-AldrichC3867-1VL55 μg/ml
Acetic acidThermoFisher Scientific103049800.02 N
DMEM/F12 with L-glutamineThermoFisher Scientific, Invitrogen11320–074n/a
DispaseThermoFisher Scientific, Invitrogen1710504124 U/ml
FBSSigma-AldrichF966510%
Penicillin/StreptomycinThermoFisher Scientific, Invitrogen15140–122100 U/ml (Pen)
100 μg/ml (Strep)
DNase IQiagen, UK7925410 μg/ml
Trypsin (from porcine pancreas)Sigma-AldrichT47990.1%
Table 4

Human Bronchiolar differentiation

https://doi.org/10.7554/eLife.26575.039
ReagentCompanyCat noFinal concentration
PneumaCultTM-ALI mediumStem Cell Technologies05001n/a
MatrigelCorning/SLS356231undiluted
Table 5

Human Alveolar differentiation

https://doi.org/10.7554/eLife.26575.040
ReagentCompanyCat noFinal concentration
Advanced DMEMThermoFisher Scientific, Invitrogen12634–010n/a
Penicillin/StreptomycinThermoFisher Scientific, Invitrogen15140–122100 U/ml (Pen) 100 μg/ml (Strep)
HepesThermoFisher Scientific, Invitrogen15630–05610 mM
GlutamaxThermoFisher Scientific, Invitrogen35050–0382 mM
N2ThermoFisher Scientific, Invitrogen17502–0481:100
B27 (-Vit A)ThermoFisher Scientific, Invitrogen12587–0101:50
N-acetylcysteineSigma-AldrichA91651.25 mM
MatrigelCorning/SLS356231Undiluted
CHIR 99021Stem Cell Institute, University of Cambridgen/a1 μM or 3 μM
FGF10R and D345-FG-025100 ng/ml
FGF7Peprotech100–19100 ng/ml
DexamethasoneSigma-AldrichD4902-25MG50 nM
cAMPSigma-AldrichB5386-5MG0.1 mM
IBMXSigma-AldrichI5879-100MG0.1 mM
DAPTSigma-AldrichD5942-25MG50 μM
Tri-iodothyronine (T3)Sigma-AldrichT6397-100MG6.7 ng/ml
Human IGF-1R and D Systems291-G1-2001 μg/ml
Interleukin-6R and D Systems206-IL-0101 μg/ml
Table 6

Primary antibodies

https://doi.org/10.7554/eLife.26575.041
AntibodyCompanyCat noHost speciesFinal dilutionAntigen retrieval (Cryo)Antigen retrieval (Paraffin)Research Resource Identifier (RRID)
ABCA3Seven Hills Bioreagents, Cincinnati, OHWRAB-ABCA3Rabbit1:500NoYesRRID:AB_577286
Acetylated tubulin (ACT)SigmaT7451, clone 6-11B-1Mouse1:3000NoNoRRID:AB_609894
AQP5Santa Cruz Biotechnology, Dallas, TexasSc9890, clone G19Goat1:100YesYesRRID:AB_2059877
β3-TUBULINBiolegend (Covance), UKPRB-435PRabbit1:1000NoN/ARRID:AB_291637
CD90Novus Biologicals, UKNBP2-37330 (clone 7E1B11)Mouse1:200NoN/ARRID:AB_2665376
E-CADHERINThermoFisher Scientific Invitrogen13–1900Rat1:3000NoN/ARRID:AB_2533005
E-CADHERINBD Biosciences, UK610182Mouse1:500NoYes (citrate)RRID:AB_397581
ETV5Santa Cruz BiotechnologySc-22807Rabbit1:200YesN/ARRID:AB_2101008
FGFR2Santa Cruz BiotechnologySC-122Rabbit1:200NoN/ARRID:AB_631509
FOXA2Santa Cruz BiotechnologySC-6554Goat1:200NoN/ARRID:AB_2262810
FOXJ1Thermo Fisher Scientific14-9965-82Mouse1:200YesYes (citrate); needs streptavidin-biotin amplificationRRID:AB_1548835
HMGA1BR and D Systems, UKAF5956Sheep1:50YesN/ARRID:AB_1964602
HMGA2Proteintech, UK20795–1-APRabbit1:100NoN/ARRID:AB_2665377
HNF1BProteintech12533–1-APRabbit1:100YesN/ARRID:AB_2116758
HOPXSanta Cruz BiotechnologySC-30216Rabbit1:50NoYes (citrate)RRID:AB_2120833
HTI-56Gift from Leland Dobbsn/aMouse1:100NoN/ARRID:AB_2665380
HTII-280Gift from Leland Dobbsn/aMouse IgM1:100NoNoRRID:AB_2665381
Human Nuclei (HuNu)Merck, UKMAB1281Mouse1:3000No (needs streptavidin-biotin amplification)N/ARRID:AB_11212527
ID2Abcam, UKAb52093Rabbit1:200YesN/ARRID:AB_880731
KRT5CovancePRB-160P-100Rabbit1:500NoYes (citrate)RRID:AB_291581
KI67BD Transduction Laboratories, UK550609, clone B56Mouse1:100NoYes (citrate)RRID:AB_393778
LPCAT1Proteintech16112–1-APRabbit1:500NoYes (citrate)RRID:AB_2135554
MUC5ACThermoFisher ScientificMS-145PMouse1:500NoYes (citrate)RRID:AB_62731
NKX2-1AbcamAb76013Rabbit1:500YesYes (citrate)RRID: AB_1310784
PDGFRACell Signalling3174 (clone D1E1E)Rabbit1:1000NoN/ARRID:AB_2162345
PDGFRBCell Signalling3169 (clone 28E1)Rabbit1:100NoN/ARRID:AB_2162497
PDPNProteintech11629–1-APRabbit1:200NoYesRRID:AB_2162067
PDPNR and D SystemsAF3670Sheep1:200NoYesRRID:AB_2162070
PECAM (CD31)AbcamAb9498Mouse1:200NoN/ARRID:AB_307284
SCGB1ASanta Cruz BiotechnologySC-25555Rabbit1:200NoYes (citrate or trypsin)RRID:AB_2269914
pro-SFTPCMillipore, UKAb3786Rabbit1:500NoYes (citrate)RRID:AB_91588
SMASigmaA5228, clone 1A4Mouse1:500NoN/ARRID:AB_262054
SOX2Santa Cruz BiotechnologySC-17320Goat1:250No/YesYes (citrate)RRID:AB_2286684
SOX9Santa Cruz BiotechnologySC-20095Rabbit1:200NoYes (citrate)RRID:AB_661282
SOX9Abcamab196450Rabbit1:200NoN/ARRID:AB_2665383
SPRY2Abcamab50317Rabbit1:200YesN/ARRID:AB_882688
TP63Cell Signaling13109Rabbit1:200YesYes (citrate); needs streptavidin-biotin amplificationRRID:AB_2637091
VECADR and D SystemsAF938Goat1:400NoN/ARRID:AB_355726
Table 7

Secondary antibodies

https://doi.org/10.7554/eLife.26575.042
AntibodyCompanyCat noFinal dilutionResearch Resource Identifier (RRID)
Donkey α-mouse 488Thermo Fisher ScientificA212021:2000RRID:AB_141607
Donkey α-rabbit 488Thermo Fisher ScientificA212061:2000RRID:AB_2535792
Donkey α-rat 488Thermo Fisher ScientificA212081:2000RRID:AB_2535794
Donkey α-mouse 546Thermo Fisher ScientificA100361:2000RRID:AB_2534012
Donkey α-rabbit 546Thermo Fisher ScientificA100401:2000RRID:AB_2534016
Donkey α-goat 555Thermo Fisher ScientificA214321:2000RRID:AB_2535853
Donkey α-rat 594Thermo Fisher ScientificA212091:2000RRID:AB_2535795
Donkey α-sheep 594Jackson Immunoresearch, West Grove, PA713-585-1471:2000RRID:AB_2340748
Donkey α-mouse 647Thermo Fisher ScientificA315711:2000RRID:AB_162542
Donkey α-rabbit 647Thermo Fisher ScientificA315731:2000RRID:AB_2536183
Goat α-rat 647Thermo Fisher ScientificA212471:2000RRID:AB_141778
Streptavidin 594Jackson Immunoresearch016-580-0841.8 μg/mlRRID:AB_2337250
Biotin-SP-conjugated Donkey α-mouseJackson Immunoresearch715-065-1501:500RRID:AB_2307438
Table 8

RT-PCR primers

https://doi.org/10.7554/eLife.26575.043
TaqMan primerCompanyCat noId no
GAPDHThermo Fisher Scientific# 4326317En/a
SOX2Thermo Fisher Scientific# 4331182Hs01053049_s1
SOX9Thermo Fisher Scientific# 4331182Hs01001343_g1
TBX4Thermo Fisher Scientific# 4331182Hs00218515_m1
Table 9

Published whole foetal lung RNAseq (Bernstein et al., 2010).

https://doi.org/10.7554/eLife.26575.044
Sample labelDescriptionSourceDonor IDAccession #
Lung_A_6Foetal day 105GEO DatasetsH-24005GSM1101693
Lung_F_2Foetal day 105GEO DatasetsH-24111GSM1101708
Lung_F_3Foetal day 108GEO DatasetsH-23887GSM1101684
Lung_F_4Foetal day 91GEO DatasetsH-23914GSM1101685
Lung_F_5Foetal day 96GEO DatasetsH-24089GSM1101699
Lung_F_6Foetal day 98GEO DatasetsH-23964GSM1101687

Additional files

Supplementary file 1

Transcription factors and signalling molecules differentially expressed in human tip versus stalk (related to Figure 2F–L).

https://doi.org/10.7554/eLife.26575.045
Supplementary file 2

Transcription Factors with mouse orthologues upregulated > 2 fold in human tips or stalks: evidence for mouse expression pattern (related to Figure 2J and L).

https://doi.org/10.7554/eLife.26575.046

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