In-vitro experiments on living cells, toxicology studies
The goal is to compare and validate the synthetic lipid models to biologically relevant cell types and to investigate how different nano-objects interact with representative lipid single and bilayers compared to human cells with and without therapeutic incorporation.
Tasks and methodology
- Cellular uptake of fluorescence-labelled nano-complexes will be checked by Delta Vision High Resolution microscopy and with an accompanied immunofluorescence counter-staining for intracellular organelles to identify distinct cellular endocytotic routes such as clathrin- or caveolae-dependent mechanisms or intracellular dispositions of the nanoparticles.
- Nanoparticles will be applied to the upper side of the transwell-membrane where either epithelial cells are seeded on the upper side and endothelial cells on the lower side or vice versa to mimic an nanoparticle application either via inhalation or injection.
- Subsequently, nanoparticle uptake and transport will be monitored via immunofluorescence in the monolayer of the opposite side of NP-exposure to verify transport events across the barrier.
Image of primary human fibroblasts co-cultured with endothelial cells. The red fluorescence represents the expression of vWF, an endothelial cell marker. Nuclei were stained with DAPI (blue).