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An Enzyme- and Serum-free Neural Stem Cell Culture Model for EMT Investigation Suited for Drug Discovery

Sailer, Martin H. and Sarvepalli, Durga and Bregere, Catherine and Fisch, Urs and Guentchev, Marin and Weller, Michael and Guzman, Raphael and Bettler, Bernhard and Ghosh, Arkasubhra and Hutter, Gregor. (2016) An Enzyme- and Serum-free Neural Stem Cell Culture Model for EMT Investigation Suited for Drug Discovery. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS (114). p. 54018.

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Official URL: https://edoc.unibas.ch/61405/

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Abstract

Epithelial to mesenchymal transition (EMT) describes the process of epithelium transdifferentiating into mesenchyme. EMT is a fundamental process during embryonic development that also commonly occurs in glioblastoma, the most frequent malignant brain tumor. EMT has also been observed in multiple carcinomas outside the brain including breast cancer, lung cancer, colon cancer, gastric cancer. EMT is centrally linked to malignancy by promoting migration, invasion and metastasis formation. The mechanisms of EMT induction are not fully understood. Here we describe an in vitro system for standardized isolation of cortical neural stem cells (NSCs) and subsequent EMT-induction. This system provides the flexibility to use either single cells or explant culture. In this system, rat or mouse embryonic forebrain NSCs are cultured in a defined medium, devoid of serum and enzymes. The NSCs expressed Olig2 and Sox10, two transcription factors observed in oligodendrocyte precursor cells (OPCs). Using this system, interactions between FGF-, BMP- and TGFbeta-signaling involving Zeb1, Zeb2, and Twist2 were observed where TGFbeta-activation significantly enhanced cell migration, suggesting a synergistic BMP-/TGFbeta-interaction. The results point to a network of FGF-, BMP- and TGFbeta-signaling to be involved in EMT induction and maintenance. This model system is relevant to investigate EMT in vitro. It is cost-efficient and shows high reproducibility. It also allows for the comparison of different compounds with respect to their migration responses (quantitative distance measurement), and high-throughput screening of compounds to inhibit or enhance EMT (qualitative measurement). The model is therefore well suited to test drug libraries for substances affecting EMT.
Faculties and Departments:03 Faculty of Medicine > Bereich Operative Fächer (Klinik) > Kopfbereich > Neurochirurgie (Guzman)
03 Faculty of Medicine > Departement Klinische Forschung > Bereich Operative Fächer (Klinik) > Kopfbereich > Neurochirurgie (Guzman)
03 Faculty of Medicine > Departement Biomedizin > Department of Biomedicine, University Hospital Basel > Brain Ischemia and Regeneration (Guzman)
03 Faculty of Medicine > Departement Biomedizin > Division of Physiology > Molecular Neurobiology Synaptic Plasticity (Bettler)
UniBasel Contributors:Bettler, Bernhard and Guzman, Raphael
Item Type:Article, refereed
Article Subtype:Research Article
ISSN:1940-087X (Electronic)1940-087X (Linking)
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:19 Jan 2019 18:37
Deposited On:19 Jan 2019 18:37

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