title: An Enzyme- and Serum-free Neural Stem Cell Culture Model for EMT Investigation Suited for Drug Discovery.
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 TGFβ-signaling involving Zeb1, Zeb2, and Twist2 were observed where TGFβ-activation significantly enhanced cell migration, suggesting a synergistic BMP-/TGFβ-interaction. The results point to a network of FGF-, BMP- and TGFβ-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.
authors: Dept. of Biomedicine, Pharmacenter, University of Basel; martin.sailer@unibas.ch.
authors: Martin H M
authors: MH
authors: Sailer
authors: Molecular Signalling and Gene Therapy, Narayana Nethralaya Foundation, Narayana Health City.
authors: Durga
authors: D
authors: Sarvepalli
authors: Brain Ischemia and Regeneration, Department of Biomedicine, University Hospital Basel.
authors: Catherine
authors: C
authors: Brégère
authors: Brain Ischemia and Regeneration, Department of Biomedicine, University Hospital Basel.
authors: Urs
authors: U
authors: Fisch
authors: Department of Neurosurgery, Klinikum Idar-Oberstein.
authors: Marin
authors: M
authors: Guentchev
authors: Department of Neurology, Laboratory of Molecular Neuro Oncology, University Hospital of Zurich.
authors: Michael
authors: M
authors: Weller
authors: Brain Ischemia and Regeneration, Department of Biomedicine, University Hospital Basel.
authors: Raphael
authors: R
authors: Guzman
authors: Dept. of Biomedicine, Pharmacenter, University of Basel.
authors: Bernhard
authors: B
authors: Bettler
authors: Molecular Signalling and Gene Therapy, Narayana Nethralaya Foundation, Narayana Health City.
authors: Arkasubhra
authors: A
authors: Ghosh
authors: Department of Neurosurgery and Institute for Stem Cell Biology and Regenerative Medicine, Stanford University.
authors: Gregor
authors: G
authors: Hutter
mesh_terms: D000818:Animals; D055808:Drug Discovery; D058750:Epithelial-Mesenchymal Transition / Q000502:physiology*; D006801:Humans; D058953:Neural Stem Cells; D051381:Rats; D015203:Reproducibility of Results; D016212:Transforming Growth Factor beta
chemical_list: D016212:Transforming Growth Factor beta