Evaluation of umbilical cord mesenchymal stem cell labeling with superparamagnetic iron oxide nanoparticles coated with dextran and complexed with Poly-L-lysine

Tatiana Taís Sibov,Luciana Cavalheiro Marti,Lorena Favaro Pavon,Daniela Mara De Oliveira,Liza Aya Mabuchi Miyaki,Lionel Fernel Gamarra,Walter Humberto Cardenas,Luiz Roberto Sardinha,Javier Bustamante Mamani

doi:10.1590/s1679-45082012000200011

Tatiana Taís Sibov, Luciana Cavalheiro Marti + Show 7 more

Open Access

https://doi.org/10.1590/s1679-45082012000200011

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Abstract

The objective of this study was to evaluate the effect of the labeling of umbilical cord vein derived mesenchymal stem cells with superparamagnetic iron oxide nanoparticles coated with dextran and complexed to a non-viral transfector agent transfector poly-L-lysine. The labeling of mesenchymal stem cells was performed using the superparamagnetic iron oxide nanoparticles/dextran complexed and not complexed to poly-L-lysine. Superparamagnetic iron oxide nanoparticles/dextran was incubated with poly-L-lysine in an ultrasonic sonicator at 37°C for 10 minutes for complex formation superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine by electrostatic interaction. Then, the mesenchymal stem cells were incubated overnight with the complex superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine and superparamagnetic iron oxide nanoparticles/dextran. After the incubation period the mesenchymal stem cells were evaluated by internalization of the complex superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine and superparamagnetic iron oxide nanoparticles/dextran by Prussian Blue stain. Cellular viability of labeled mesenchymal stem cells was evaluated by cellular proliferation assay using 5,6-carboxy-fluorescein-succinimidyl ester method and apoptosis detection by Annexin V- Propidium Iodide assay. mesenchymal stem cells labeled with superparamagnetic iron oxide nanoparticles/dextran without poly-L-lysine not internalized efficiently the superparamagnetic iron oxide nanoparticles due to its low presence detected within cells. Mesenchymal stem cells labeled with the complex superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine efficiently internalized the superparamagnetic iron oxide nanoparticles due to greater presence in the cells interior. The viability and apoptosis assays demonstrated that the mesenchymal stem cells labeled and not labeled respectively with the superparamagnetic iron oxide nanoparticles/dextran/poly-L-lysine continue to proliferate over seven days and the percentage of cells in early or late apoptosis is low compared to the percentage of live cells over the three days. Our results showed that the use of poly-L-lysine complexed with superparamagnetic iron oxide nanoparticles/dextran provides better internalization of these superparamagnetic iron oxide nanoparticles in mesenchymal stem cells Thus, we demonstrated that this type of labeling is not cytotoxic to the mesenchymal stem cells, since the viability and apoptosis assays showed that the cells remain alive and proliferating. The efficiency of this type of labeling in mesenchymal stem cells can provide non-invasive methods for monitoring these cells in vivo.

Highlights

Superparamagnetic iron oxide nanoparticles (SPIONs) have been widely used in various biomedical applications in vitro and in vivo[1,2]
In the immunophenotyping assay performed by flow cytometry, the MSCs did not express positivity for CD14, CD31, CD34, CD45, CD106, CD133 and Cellular death assay – annexin/propidium iodide For the cellular death assay, the BD Pharmingen FITC Annexin V Apoptosis Detection Kit I
The MSCs expressed positivity for HLA A, B and C (HLA-I), CD29, CD73, CD90 and CD105; the results are represented by histograms and the cell populations are labeled positively; the cells labeled with control isotopes are superimposed on the same graph (Figure 2)

Summary

Introduction

Superparamagnetic iron oxide nanoparticles (SPIONs) have been widely used in various biomedical applications in vitro and in vivo[1,2] In these applications, there is the need for high specificity, efficiency and quick internalization of the SPION into specific target cells, but this may be limited by various factors[3]: (i) aggregation of SPIONs (SPIONs have a high surface/volume ratio, and tend to agglomerate); (ii) short half-life of SPIONs in blood circulation (when the SPIONs agglomerate, or adsorb plasma proteins, which are rapidly eliminated from the blood circulation by macrophages from the phagocyte mononuclear system before they can reach the target cells); (iii) and the low efficiency of intracellular capture of SPIONs. Biocompatible polymers and agents with specific targets are, bound to the surface of SPIONs to avoid agglomeration and to increase its nonspecific intracellular absorption[4,5]. The nonefficient capture of SPION-DX has been reduced by coating these SPIONs with polymers[2]

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