Smart Nano-objects for Alteration of Lipid bilayers (SNAL) Initial Training Network is a multidisciplinary EU-funded 7th Framework Programme specially designed to provide scientific and transferable skill training and career development for early stage researchers and experienced researchers in the field of lipid research.

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ESR11-F: Delivery of molecular payloads through primary human cell membrane

In-vitro experiments on living cells, toxicology studies

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Andre Dias

Objectives

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.

HF-HDMEC Co-culture vWF - Alexa Fluor 546 Phase-contrast PF

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).

Starting date October 2014

Related research topics

André Dias

MSCA Early Stage Researcher at the Universitaet Frankfurt, Germany Nanotoxicity and in-vitro investigation of interaction of nanomaterials with living cells. more...
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