Large (>100 pg/cell) amounts of internalized IONPs for specific surface coated (charged) nanoparticles have been reported, but using long incubation times (c.a. Most in vivo cell-labeling methods with IONPs are based on direct incubation (DI), which relies on standard endocytosis mechanisms and requires long periods of incubation and large IONPs concentrations 12, because ultimately the cellular internalization of IONPs depends upon the sedimentation and diffusion velocities of the nanoparticles 13. Table S3) 10, 11.Ĭurrent labeling procedures, however, present important limitations that undermine its potential, most of them related to the reduced intracellular concentration of IONPs and the time consuming labelling procedure. Thus, IONPs-labeled cells is one of the most promising approaches for a quick and reliable evaluation of grafted cells in preclinical studies and clinical trials ( Suppl. By using iron oxide nanoparticles (IONPs) as contrast agents 10, it can be done in vivo in a non-invasive manner and it has been shown to be safe and effective. Magnetic resonance imaging (MRI) offers several advantages for tracking, such as high resolution, easy accessibility and three-dimensional capabilities 6, 7, 8, 9. The cell-based therapy relies on the successful delivery of labeled cells into the target site and therefore, tracking these became a main issue to ensure clinical safety and therapeutic efficacy 5. The possibility of labeling living cells has allowed the development during the last decade of a variety of techniques within the biomedical field, ranging from in vitro cell manipulation to imaging-based applications in the clinical settings, especially relevant in cancer and cell therapy 1, 2, 3, 4. Similar trends are found in human mesenchymal stem cells, thereby demonstrating the general feasibility of the methodology, which is easily transferable to any laboratory with great potential for the development of improved biomedical applications. CMI results in controllable cellular uptake efficiencies at least three orders of magnitude larger than current procedures. Here we show that centrifugation-mediated internalization (CMI) promotes a high uptake of IONPs in glioblastoma tumour cells, just in a few minutes and via clathrin-independent endocytosis pathway. Current procedures to label living cells with IONPs are based on direct incubation or physical approaches based on magnetic or electrical fields, which always display very low cellular uptake efficiencies. Iron oxide nanoparticles (IONPs)-labelled cells is one of the most promising approaches for a fast and reliable evaluation of grafted cells in both preclinical studies and clinical trials. Nanotechnology plays an increasingly important role in the biomedical arena.
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