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Raluca Maria Fratila

Real-time monitoring of cellular effects of sublethal magnetic hyperthermia.

The research proposed for this COST STSM is part of my on-going two-years Marie Skłodowska-Curie (MSC) Project OUTstandINg, which is focused on the covalent immobilization of magnetic nanoparticles (MNPs) on living cell plasma membranes using bioorthogonal click chemistry with the aim to address two fundamental questions in the field of magnetic hyperthermia: 1) how the subcellular localization (on the plasma membrane or inside the cells) of MNPs affects their heating behaviour when compared to MNPs in solution 2) how MNPs immobilization and sub-lethal magnetic hyperthermia impact different subcellular signalling pathways and the biophysics of cell membranes. The aim of the STSM at UCL Healthcare Biomagnetics Laboratory was to explore the possibility of real-time monitoring of the effect of sublethal magnetic hyperthermia on living cells, using the in situ MHT/imaging system developed by the host group (C. Blanco-Andujar et al., Nanomedicine, 2016, 11, 121). The preliminary studies carried out during the STSM were focused on optimizing the experimental conditions in order to maintain cells healthy and functioning normally on the microscope stage for the whole duration of the experiment (usually up to 16 h). This is one of the main technical challenges for performing live-cell imaging experiments and is fundamental for our studies as the decrease of cellular viability non-associated with the magnetic hyperthermia experiment must be minimized. To this end, MCF-7 (human breast adenocarcinoma) cells were cultured on 35-mm cell culture dishes at 37 ºC in a humidified atmosphere containing 5% CO₂. Different control experiments were carried out to establish the best conditions for long-term imaging experiments. These experiments included cell growth using different cell culture dishes and use of an anti-evaporation oil to minimize changes in osmolarity due to evaporation, which can be severe when using relatively low volumes of cell culture medium in a 37 ºC environment.