Mus musculus
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA358829 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:Two populations of Nkx2-1+ progenitors in the developing foregut endoderm give rise to the entire post-natal lung and thyroid epithelium, but little is known about these cells, as they are difficult to isolate in a pure form. We demonstrate here the purification and directed differentiation of primordial lung and thyroid progenitors derived from mouse embryonic stem cells (ESCs). Inhibition of TGFM-NM-2 and BMP signaling, followed by combinatorial stimulation of BMP and FGF signaling can specify these cells efficiently from definitive endodermal precursors. When derived using Nkx2-1GFP knock-in reporter ESCs, these progenitors can be purified for expansion in culture and have a transcriptome that overlaps with developing lung epithelium. Upon induction, they can express a broad repertoire of markers indicative of lung and thyroid lineages and can recellularize a 3D lung tissue scaffold. Thus, we have derived a pure population of progenitors able to recapitulate the developmental milestones of lung/thyroid development. Comparison between Nkx2-1 positive and Nkx2-1 negative cells derived after 14 days of differentiation of mouse ES cells carrying a knock-in reporter gene, Nkx2-1GFP.
2012-04-04 | E-GEOD-35063 | biostudies-arrayexpress
Project description:Two populations of Nkx2-1+ progenitors in the developing foregut endoderm give rise to the entire post-natal lung and thyroid epithelium, but little is known about these cells, as they are difficult to isolate in a pure form. We demonstrate here the purification and directed differentiation of primordial lung and thyroid progenitors derived from mouse embryonic stem cells (ESCs). Inhibition of TGFβ and BMP signaling, followed by combinatorial stimulation of BMP and FGF signaling can specify these cells efficiently from definitive endodermal precursors. When derived using Nkx2-1GFP knock-in reporter ESCs, these progenitors can be purified for expansion in culture and have a transcriptome that overlaps with developing lung epithelium. Upon induction, they can express a broad repertoire of markers indicative of lung and thyroid lineages and can recellularize a 3D lung tissue scaffold. Thus, we have derived a pure population of progenitors able to recapitulate the developmental milestones of lung/thyroid development.
2012-04-05 | GSE35063 | GEO
Project description:The in vitro directed differentiation of pluripotent stem cells (PSCs) through stimulation of developmental signaling pathways can generate mature somatic cell types for basic laboratory studies or regenerative therapies. We used microarrays to detail the global transcriptomes of mouse embryonic stem cells differentiated in vitro into putative thyroid vs lung epithelial lineages using serum-free media supplemented with either BMP4+FGF2 (thyroid media) or BMP4+Wnt3a (lung media.) Differentiated cells carried an Nkx2-1mCherry knock-in reporter to allow sorting of mCherry + vs - cells in each condition on day 14 of differentiation for global transcriptomic profiling.
2017-10-31 | GSE92916 | GEO
Project description:It has been postulated that during human fetal development all cells of the lung epithelium derive from an embryonic endodermal NKX2-1+ precursor, however, this hypothesis has not been formally tested due to an inability to purify or track this theorized cell for detailed characterization. Here we engineer and developmentally differentiate NKX2-1GFP reporter pluripotent stem cells (PSCs) in vitro to generate and isolate a human primordial lung progenitor that expresses NKX2-1 but is initially devoid of markers of differentiated lung lineages. As these progenitors move through the earliest moments of lung lineage specification from definitive endoderm they can be imaged in real time or isolated for time-series global transcriptomic profiling. We performed microarray analysis of 5 timepoints of human iPSC to lung directed differentiation compared to week 21 human fetal lung and Neural NKX2-1+ cell controls. These profiles indicate that evolutionarily conserved, stage-dependent developmental gene signatures are expressed in primordial human lung progenitors. Using a TALEN-targeted fluorescent reporter to purify iPSC-derived lung progenitors (C17 NKX2-1GFP) we analyzed cells at major developmental time points in vitro (undifferentiated iPSC, definitive endoderm, anterior foregut endoderm and sorted NKX2-1GFP+ and NKX2-1GFP- cells on day 15 and day 28 of the protocol). We also differentiated NXK2-1GFP iPSC in a neural protocol and isolated neural NKX2-1GFP+ cells. Approximately 90% pure human fetal lung epithelial cells from week 21 embryos were used as controls.
2017-05-02 | GSE83310 | GEO
Project description:It has been postulated that during human fetal development all cells of the lung epithelium derive from an embryonic endodermal NKX2-1+ precursor, however, this hypothesis has not been formally tested due to an inability to purify or track this theorized cell for detailed characterization. Progress has been made in deriving lung epithelial cells from human induced pluripotent stem cells (iPSCs). However, little is known about the heterogeneity or genetic programs of the cells generated using these lung differentiation protocols. We engineered and differentiated NKX2-1GFP reporter iPSCs in vitro, recapitulating the major developmental milestones of lung development, to generate and isolate human primordial lung progenitors (day 15 of differentiation) that expresses NKX2-1 but are initially devoid of markers of differentiated lung lineages. To further characterize the cells generated in the lung directed differentiation protocol we performed single cell RNA-seq analysis of cells on day 15 of lung directed differentiation. We analyzed sorted NKX2-1GFP+ cells for the iPSC line C17 and cells from the iPSC line BU3.
2017-05-02 | GSE96106 | GEO
Project description:This study provides ultra-deep bulk RNA-seq data from human embryonic stem cell (hESC)-derived thyroid follicular cells. The NKX2-1GFP+ population was isolated using fluorescence-activated cell sorting (FACS) and subjected to RNA extraction. Library preparation was performed using the Ovation Solo RNA-seq System, followed by high-output sequencing on the Illumina HiSeq 1500 platform. Each sample generated approximately 400 million reads. These data enable transcriptomic analysis of differentiated thyroid lineage cells exposed to IFN-α and IFN-γ.
2025-08-13 | GSE295120 | GEO
Project description:RNA-seq analyses of MEFs obtained from GFP/SP-C transgenic mice on CBA/Ca x C57BL/6 mixed background were transduced sets of 3 or 4 combination factors (Nkx2-1/Foxa1/Gata, Nkx2-1/Foxa2/Gata, Nkx2-1/Foxa1/Foxa2/Gata6) into induced pulmonary epithelial cell-like cells 14 days after the transduction. Abstract is as follows: pneumocyte differentiation has been achieved previously using embryonic stem cells or induced pluripotent stem cells. However, direct reprograming of somatic cells into pulmonary epithelial cells has not been achieved. Here, we report that a combination of three or four transcription factors (i.e., Nkx2-1, Gata6, and [Foxa1 and/or Foxa2]) directly reprogrammed mouse tail-tip fibroblast or embryonic fibroblast into differentiated induced pulmonary epithelial cell-like cells (iPUL cells). iPUL cells had a global gene expression profile similar to various kinds of pulmonary epithelial cells. Some iPUL cells showed lamellar body-like structures and expressed SP-C, consistent with the features of alveolar epithelial type II cells. Interestingly, intratracheal administration of iPUL cells rescued influenza virus-induced acute lung injury in mice. These findings demonstrate that functional pulmonary epithelial cell-like cells can be directly reprogrammed from differentiated somatic cells by defined factors. Reprograming of fibroblasts might provide a source of pulmonary epithelial cells for regenerative medicine. Publicly available raw RNA-seq data (GSE80101) representing whole mouse lung (SRR3353494, SRR3353497, SRR3353498) and alveolar type II cells (SRR3353495, SRR3353500, SRR3353510, SRR3353512) sorted in the basis of EpCAM+ and dTomato+ from SpcCreERT2;RosatdTomato mice were also simultaneously processed with our data.
2023-06-30 | GSE109810 | GEO
Project description:Co-cultures of lung epithelium and mesenchyme are useful tools to study epithelial-mesenchymal crosstalk in lung development and disease. However, many previous attempts to generate such co-cultures have yielded poor juxtaposition between the epithelial and the mesenchymal lineage. In addition, induced pluripotent stem cell (iPSC)-derived co-cultures often contain generic mesenchyme that is not necessarily lung-specific. We sought to establish co-cultures of purified mouse iPSC-derived lung-specific mesenchyme and iPSC-derived lung epithelial progenitors. We used a mouse iPSC line carrying a lung mesenchyme-specific reporter/tracer (Tbx4-LERGFP) to generate lung mesenchymal progenitors by directed differentiation via a lateral plate mesodermal progenitor state (induced lung mesenchyme, iLM). In parallel we differentiated a mouse embryonic stem (ES) cell line carrying a Nkx2-1mCherry reporter into lung epithelial progenitor cells using our established directed differentiation protocol. We then combined the purified lung epithelial and mesenchymal progenitor cells and co-cultured them in distal or proximal differentiation media for 1 week on Matrigel. We find that cells self-organize into complex 3-dimensional organoids with closely juxtaposed epithelial and mesenchymal cells. Furthermore, co-culture affects the molecular phenotype of both lineages. Our iPSC-derived co-culture model can provide an inexhaustible source of cells for studying lung development, modeling diseases, and developing therapeutics.
2023-05-08 | GSE228313 | GEO
Project description:Multipotent Nkx2-1-positive lung epithelial primordial progenitors of the foregut endoderm are thought to be the developmental precursors to all adult lung epithelial lineages. However, little is known about the global transcriptomic programs or gene networks that regulate these gateway progenitors in vivo due to their rarity and transient presence during a narrow developmental window, embryonic day E9.0 in mice. Here we describe the unique genetic program of in vivo lung primordial progenitors and computationally identify the signaling pathways that are involved in their cell-fate determination from pre-specified embryonic foregut. We integrate this information in computational models to generate in vitro engineered lung primordial progenitors from mouse pluripotent stem cells, improving the fidelity of the resulting cells through unbiased, easy-to-interpret similarity scores and modulation of cell culture conditions, including biomechanical cues. As the genetic characterization of early in vivo embryonic progenitors is rapidly expanding, the methodology proposed here can have wide applicability to the in vitro derivation of bona fide tissue progenitors of all germ layers.
2019-12-20 | GSE138904 | GEO
Project description:Multipotent Nkx2-1-positive lung epithelial primordial progenitors of the foregut endoderm are thought to be the developmental precursors to all adult lung epithelial lineages. However, little is known about the global transcriptomic programs or gene networks that regulate these gateway progenitors in vivo due to their rarity and transient presence during a narrow developmental window, embryonic day E9.0 in mice. Here we describe the unique genetic program of in vivo lung primordial progenitors and computationally identify the signaling pathways that are involved in their cell-fate determination from pre-specified embryonic foregut. We integrate this information in computational models to generate in vitro engineered lung primordial progenitors from mouse pluripotent stem cells, improving the fidelity of the resulting cells through unbiased, easy-to-interpret similarity scores and modulation of cell culture conditions, including biomechanical cues. As the genetic characterization of early in vivo embryonic progenitors is rapidly expanding, the methodology proposed here can have wide applicability to the in vitro derivation of bona fide tissue progenitors of all germ layers.
2019-12-20 | GSE139186 | GEO