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European Biophysics Journal

, Volume 46, Issue 5, pp 433–444 | Cite as

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

  • Vratislav Cmiel
  • Josef SkopalikEmail author
  • Katerina PolakovaEmail author
  • Jan Solar
  • Marketa Havrdova
  • David Milde
  • Ivan Justan
  • Massimiliano Magro
  • Zenon Starcuk
  • Ivo Provaznik
Original Article

Abstract

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.

Keywords

Intracellular fluorescent labels Stem cell tracking Dual contrast agents Iron oxide nanoparticles Confocal microscopy Mesenchymal stromal cells Rhodamine 

Notes

Acknowledgements

The authors acknowledge the support by Ministry of Education, Youth and Sports  of the Czech Republic (project LO1305), the Operational Program “Education for Competitiveness – European Social Fund” CZ.1.07/2.3.00/20.0155. This work was also supported by the project no. LQ1605 from the National Program of Sustainability II (MEYS CR) and by the project FNUSA-ICRC no. CZ.1.05/1.1.00/02.0123 (OP VaVpI). Support by Ministry of Education of the Czech Republic (LO1212 and LO1305, CZ.1.05/2.1.00/01.0017) and the support by Student Project IGA­PrF­2015­017 is also acknowledged.

Supplementary material

249_2016_1187_MOESM1_ESM.pdf (1 mb)
Supplementary material 1 (PDF 1072 kb)

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Copyright information

© European Biophysical Societies' Association 2016

Authors and Affiliations

  • Vratislav Cmiel
    • 1
  • Josef Skopalik
    • 2
    Email author
  • Katerina Polakova
    • 3
    Email author
  • Jan Solar
    • 4
  • Marketa Havrdova
    • 3
  • David Milde
    • 3
  • Ivan Justan
    • 2
    • 4
  • Massimiliano Magro
    • 5
  • Zenon Starcuk
    • 6
  • Ivo Provaznik
    • 1
  1. 1.Department of Biomedical EngineeringFEEC, Brno University of TechnologyBrnoCzech Republic
  2. 2.Department of Human Pharmacology and Toxicology, Faculty of PharmacyUniversity of Veterinary and Pharmaceutical SciencesBrnoCzech Republic
  3. 3.Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Experimental Physics and Analytical Chemistry, Faculty of SciencePalacky UniversityOlomoucCzech Republic
  4. 4.International Clinical Research CenterSt. Anne’s University Hospital BrnoBrnoCzech Republic
  5. 5.Department of Comparative Biomedicine and Food ScienceUniversity of PaduaPaduaItaly
  6. 6.Institute of Scientific Instruments, The Czech Academy of SciencesBrnoCzech Republic

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