The invention relates to the method for in vitro loading of biological eukaryotic cells with polymer coated magnetic nanoparticles that enables high loading of nanoparticles into biological cells where the cells retain high viability and where the loaded nanoparticles exhibit multimodal function involving optical and transmission electron imaging, and allow external manipulation with magnetic field. The method describes protocol for in vitro loading with nanoparticles having magnetic core and have coating with at least one or several polymer layers. The protocol and developed nanoparticles enable high loading of NPs inside the cell with small effect on cellular viability and can be thus used for cell labeling and tracking in different biomedical and biotechnological applications like tissue regeneration or tracking of stem cells.
F.32 International patent
COBISS.SI-ID: 10825556Magnetic nanoparticles are already used in biotechnological applications. Coating of magnetic nanoparticles is crucial for their stability in biological environments. In this work we tested the effect of polyethyleneimine coated magnetic nanoparticles (PEI NP) on CHO cell line. We analysed if additional coating with L-Glutathione Reduced (PEI-glutathione NP) can reduce cytotoxicity of PEI-NPs. First, we synthesized PEI NP and PEI-glutathione NP and second, characterization with Dynamic Light Scattering and zeta potential measurements was made. DLS and zeta potential measurements confirmed additional L-Glutathione coating. Viability was determined with fluorescent staining; metabolic activity was measured with MTS assay. Also presence of reactive oxygen species was measure while transmission electron microscopy was used for visualization of internalization of different nanoparticles. Our results show, that additional L-Glutathione coating reduces cytotoxic effect of analysed NPs on CHO cells. Toxicity might have been reduced as a result of L-glutathione antioxidant function or as a result of lower zeta potential. In this study we have shown that reactive oxygen species plays an important role in PEI NP cytotoxicity. TEM analysis confirmed that PEI NP and PEI-glutathione NP enter the cell via macropinocytosis, PEI-glutathione NP also via clathrin-mediated endocytosis.
F.04 Increase of the technological level
COBISS.SI-ID: 8023929