title: Rhodamine bound maghemite as a long-term dual imaging nanoprobe of adipose tissue-derived mesenchymal stromal cells.
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.
authors.affiliation: Department of Biomedical Engineering, FEEC, Brno University of Technology, Brno, Czech Republic.
authors.forename: Vratislav
authors.initials: V
authors.lastname: Cmiel
authors.affiliation: Department of Human Pharmacology and Toxicology, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic. j.skopalik@gmail.com.
authors.forename: Josef
authors.initials: J
authors.lastname: Skopalik
authors.affiliation: Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Experimental Physics and Analytical Chemistry, Faculty of Science, Palacky University, Olomouc, Czech Republic. dr.kacka.polakova@gmail.com.
authors.forename: Katerina
authors.initials: K
authors.lastname: Polakova
authors.affiliation: International Clinical Research Center, St. Anne's University Hospital Brno, Brno, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Solar
authors.affiliation: Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Experimental Physics and Analytical Chemistry, Faculty of Science, Palacky University, Olomouc, Czech Republic.
authors.forename: Marketa
authors.initials: M
authors.lastname: Havrdova
authors.affiliation: Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry, Experimental Physics and Analytical Chemistry, Faculty of Science, Palacky University, Olomouc, Czech Republic.
authors.forename: David
authors.initials: D
authors.lastname: Milde
authors.affiliation: Department of Human Pharmacology and Toxicology, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic.|International Clinical Research Center, St. Anne's University Hospital Brno, Brno, Czech Republic.
authors.forename: Ivan
authors.initials: I
authors.lastname: Justan
authors.affiliation: Department of Comparative Biomedicine and Food Science, University of Padua, Padua, Italy.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Magro
authors.affiliation: Institute of Scientific Instruments, The Czech Academy of Sciences, Brno, Czech Republic.
authors.forename: Zenon
authors.initials: Z
authors.lastname: Starcuk
authors.affiliation: Department of Biomedical Engineering, FEEC, Brno University of Technology, Brno, Czech Republic.
authors.forename: Ivo
authors.initials: I
authors.lastname: Provaznik
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D002470:Cell Survival; D003911:Dextrans / Q000378:metabolism; D005260:Female; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D057054:Molecular Imaging / Q000379:methods*; D015335:Molecular Probes / Q000737:chemistry* / Q000378:metabolism; D012235:Rhodamines / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
chemical_list: D003911:Dextrans; D058185:Magnetite Nanoparticles; D015335:Molecular Probes; D012235:Rhodamines; C065507:ferumoxides
keywords: Confocal microscopy; Dual contrast agents; Intracellular fluorescent labels; Iron oxide nanoparticles; Mesenchymal stromal cells; Rhodamine; Stem cell tracking