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Generation of stem cell derived human lung epithelium in High-Throughput-Screenings

Generation of stem cell derived human lung epithelium in High-Throughput-Screenings

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BLUHMKI, Teresa, 2022. Generation of stem cell derived human lung epithelium in High-Throughput-Screenings [Dissertation]. Konstanz: University of Konstanz

@phdthesis{Bluhmki2022Gener-57114, title={Generation of stem cell derived human lung epithelium in High-Throughput-Screenings}, year={2022}, author={Bluhmki, Teresa}, address={Konstanz}, school={Universität Konstanz} }

<rdf:RDF xmlns:dcterms="http://purl.org/dc/terms/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#" xmlns:foaf="http://xmlns.com/foaf/0.1/" xmlns:void="http://rdfs.org/ns/void#" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/rdf/resource/123456789/57114"> <dcterms:rights rdf:resource="https://rightsstatements.org/page/InC/1.0/"/> <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/rdf/resource/123456789/28"/> <dcterms:issued>2022</dcterms:issued> <dcterms:abstract xml:lang="eng">In drug discovery, there is an increasing demand for more physiological in vitro models that recapitulate the disease situation in patients. Many different preclinical approaches to model lung diseases have been established over the years, but they lack throughput and the access to disease relevant cells. These central obstacles need to be overcome by the development of highly physiological primary and stem cell derived in vitro model systems that increase human relevance and throughput for therapeutic compound screenings and drug development in lung research. Paper #1 focused on the development of a miniaturized 96-Transwell microplate system where primary human small airway epithelial (hSAE) cells were cultured at an air-liquid interface (ALI). After four weeks of ALI culture, a pseudostratified epithelium containing basal, club, goblet and ciliated cells was produced. A novel custom-made device for 96-parallelized transepithelial electric resistance (TEER) measurements, confirmed the development of a tight barrier function during ALI maturation. Additionally, the miniaturized hSAE cell cultures were responsive to pro-fibrotic stimuli in a physiological manner regarding epithelial dysfunction in Idiopathic Pulmonary Fibrosis (IPF). In paper #2 a large-scale expansion of human induced pluripotent stem cells (hiPSCs) for high-throughput adaption is presented and an optimized step-wise differentiation of hiPSCs into Definitive Endoderm (DE) cells is described. The produced DE cells were demonstrated to express classical DE markers on the gene expression and protein level. The here described DE cells are multipotent progenitors and act as starting points for subsequent differentiation approaches towards alveolar-like cells. Paper #3 demonstrates a unique, miniaturized 96-Transwell microplate system where hiPSCderived alveolar-like cells were cultured under ALI conditions. To this end, hiPSCs were differentiated into lung epithelial progenitor cells (LPCs) and subsequently matured into a functional alveolar type 2 (AT2)-like epithelium with monolayer-like morphology. AT2-like cells displayed characteristics such as alveolar surfactant protein expressions and lamellar-body like structures. For the induction of a disease-like phenotype in vitro, the functional AT2-like cells were stimulated with cytokines and growth factors present in the alveolar tissue of IPF patients, leading to the production of pro-fibrotic mediators both on the mRNA and protein level. Thus, the miniaturized primary hSAE and hiPSC-derived cellular model systems enable the recapitulation of a physiologically responsive lung epithelium, while paving the route towards a miniaturized medium throughput approach of pharmaceutical drug discovery, in respect of fibrotic lung diseases.</dcterms:abstract> <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/57114"/> <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/> <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/57114/3/Bluhmki_2-1rc72gjll19g36.pdf"/> <dc:creator>Bluhmki, Teresa</dc:creator> <dc:contributor>Bluhmki, Teresa</dc:contributor> <dc:language>eng</dc:language> <foaf:homepage rdf:resource="http://localhost:8080/jspui"/> <dc:rights>terms-of-use</dc:rights> <dcterms:title>Generation of stem cell derived human lung epithelium in High-Throughput-Screenings</dcterms:title> <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/rdf/resource/123456789/28"/> <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/57114/3/Bluhmki_2-1rc72gjll19g36.pdf"/> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2022-03-31T10:15:58Z</dcterms:available> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2022-03-31T10:15:58Z</dc:date> </rdf:Description> </rdf:RDF>

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