Abstract
Adipose tissue-derived stromal cells (ASCs) represent a capable source for cell-based therapeutic approaches. For monitoring a cell-based application in vivo, magnetic resonance imaging (MRI) of cells labeled with iron oxide particles is a common method. It is the aim of the present study to analyze potential DNA damage, cytotoxicity and impairment of functional properties of human (h)ASCs after labeling with citrate-coated very small superparamagnetic iron oxide particles (VSOPs). Cytotoxic as well as genotoxic effects of the labeling procedure were measured in labeled and unlabeled hASCs using the MTT assay, comet assay and chromosomal aberration test. Trilineage differentiation was performed to evaluate an impairment of the differentiation potential due to the particles. Proliferation as well as migration capability were analyzed after the labeling procedure. Furthermore, the labeling of the hASCs was confirmed by Prussian blue staining, transmission electron microscopy (TEM) and high-resolution MRI. Below the concentration of 0.6 mM, which was used for the procedure, no evidence of genotoxic effects was found. At 0.6 mM, 1 mM as well as 1.5 mM, an increase in the number of chromosomal aberrations was determined. Cytotoxic effects were not observed at any concentration. Proliferation, migration capability and differentiation potential were also not affected by the procedure. Labeling with VSOPs is a useful labeling method for hASCs that does not affect their proliferation, migration and differentiation potential. Despite the absence of cytotoxicity, however, indications of genotoxic effects have been demonstrated.
Highlights
Blue staining clearly identified intracellular very small superparamagnetic iron oxide particles (VSOPs) within the labeled Human Adipose Tissue-Derived Stromal Cells (hASCs) (Figure 1A), whereas no positive staining could be found in unlabeled cells (Figure 1B)
After three weeks of treatment with osteogenic differentiation medium, VSOPs-labeled hASCs and untreated controls using Alizarin Red and von Kossa staining revealed a deposition of calcified extracellular matrix, which could not be detected in the negative controls
HASCs were labeled with citrate-coated VSOPs
Summary
Iron oxide nanoparticles (IONPs) are used for labeling various cell types: ASCs, mesenchymal stem cells from the bone marrow (BMSCs), chondrocytes [19,21,23,30,31], human adult neural stem cells (haNCSs) [31], murine embryonic stem cells (mESCs) [31] and human umbilical blood mononuclear cells (hUCBCs) [27] They are applied in cancer imaging and treatment and used for targeted drug and gene delivery [24,29,32,33,34]. The results should provide additional information on the interaction between VSOPs commonly used for cell labeling and hASCs
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