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In vitro studies of BDE-99 induced neurotoxicity
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
2013 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Neurospheres (NSph) are in vitro culture condition specific aggregates of neural stem and precursor stem cell (NSPC) isolated from the developing and/or adult central nervous system organisms. The development of the NSPC and NSph methodology together with high-throughput analysis methods has opened the door for a more systematic screening, hazard priority ranking, and analysis of potential developmental neurotoxicity agents. The purpose of this project was to establish and develop the NSph assay and use high content image analysis (HCA) of neuronal and synaptic growth. Neurotoxicity aspects were explored by applying the environmental pollutant BDE-99 to primary cells from rats and NSPCs from rats and mice. Neurotoxic sensitivity differences between non-dissociated and dissociated NSph cells and relation between cell density and dose response and exposure time was investigated.

BDE-99 did not disturb mouse non-dissociated NSph proliferation or migration, but reduced the total number of cells in dissociated NSph cells. A HCA comparison demonstrated furthermore that short-term cultures were more sensitive to BDE-99 exposure than long-term cultures. These results indicate that rodent proliferation and migration tests need to be developed further and that initial cell density, cell culture duration, substance concentration, time of exposure and exposure time are important factors when using HCA on dissociated NSph cells. Image analysis of the data was also a factor. Short-term and long-term plates were analyzed using the same software settings but the program could not detect neurite outgrowth in the long-term plates as it did in the short-term plates. However, collectively, these studies suggest that the NSph assay and HCA analysis are promising tools for developmental neurotoxicity testing in vitro.

Place, publisher, year, edition, pages
2013. , 24 p.
Keyword [en]
In vitro, neurosphere, BDE-99, high throughput screening, high content image analysis, developmental neurotoxicity, EGF, FGF2
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
URN: urn:nbn:se:uu:diva-202430OAI: oai:DiVA.org:uu-202430DiVA: diva2:631947
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Available from: 2013-06-24 Created: 2013-06-24 Last updated: 2013-06-24Bibliographically approved

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