Rhodamine bound maghemite as a long-term dual imaging nanoprobe of adipose tissue-derived mesenchymal stromal cells

Authors

CMIEL Vratislav SKOPALIK Josef POLAKOVA Katerina SOLAR Jan HAVRDOVA Marketa MILDE David JUSTAN Ivan MAGRO Masimiliano STARCUK Zenon PROVAZNIK Ivo

Year of publication 2016
Type Article in Periodical
Magazine / Source European Biophysics Journal
MU Faculty or unit

Faculty of Medicine

Citation
Web http://dx.doi.org/10.1007/s00249-016-1187-1
Doi http://dx.doi.org/10.1007/s00249-016-1187-1
Field Biophysics
Keywords Confocal microscopy; Dual contrast agents; Intracellular fluorescent labels; Iron oxide nanoparticles; Mesenchymal stromal cells; Rhodamine; Stem cell tracking
Description In the last few years, magnetically labeled cells have been intensively explored, and non-invasive cell tracking and magnetic manipulation methods have been tested in preclinical studies focused on cell transplantation. For clinical applications, it is desirable to know the intracellular pathway of nanoparticles, which can predict their biocompatibility with cells and the long-term imaging properties of labeled cells. Here, we quantified labeling efficiency, localization, and fluorescence properties of Rhodamine derivatized superparamagnetic maghemite nanoparticles (SAMN-R) in mesenchymal stromal cells (MSC). We investigated the stability of SAMN-R in the intracellular space during a long culture (20 days). Analyses were based on advanced confocal microscopy accompanied by atomic absorption spectroscopy (AAS) and magnetic resonance imaging. SAMN-R displayed excellent cellular uptake (24 h of labeling), and no toxicity of SAMN-R labeling was found. 83% of SAMN-R nanoparticles were localized in lysosomes, only 4.8% were found in mitochondria, and no particles were localized in the nucleus. On the basis of the MSC fluorescence measurement every 6 days, we also quantified the continual decrease of SAMN-R fluorescence in the average single MSC during 18 days. An additional set of analyses showed that the intracellular SAMN-R signal decrease was minimally caused by fluorophore degradation or nanoparticles extraction from the cells, main reason is a cell division. The fluorescence of SAMN-R nanoparticles within the cells was detectable minimally for 20 days. These observations indicate that SAMN-R nanoparticles have a potential for application in transplantation medicine.

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