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

Abstract

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

In Vitro Evaluation of Hyperthermia Magnetic Technique Indicating the Best Strategy for Internalization of Magnetic Nanoparticles Applied in Glioblastoma Tumor Cells

“…The transfection agent PLL increased the SPION Amine (100 nm) internalization by cells as shown in Figure 4 . This strategy was also evidenced in other studies [ 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 ]. The use of transfection agent as PLL, that is a linear polymer of lysine that bears a positive charge at neutral pH, benefits the electrostatic interaction between nanoparticles and cells [ 60 ], as PLL increases the SPION Amine positive charge (increasing the zeta potential) leading to enhanced attraction with C6 cell membrane which is negatively charged.…”

Induction of neural stem cell proliferation by iron oxide nanoparticles and magnetic field and Ki67 gene expression in rat hippocampus after ischemia/reperfusion

“…Furthermore, Huang et al indicated that iron oxide nanoparticles increase the proliferation of human mesenchymal stem cells (17). Moreover, Sibov et al demonstrated that iron oxide nanoparticles along with dextran and poly-L-lysine could increase the proliferation of human umbilical cord mesenchymal stem cells while reducing the apoptosis in these cells (18). Additionally, Sadek et al reported that using mesenchymal stem cells reduced the ischemia-reperfusion injury in the kidney of the albino mice (19).…”

Easy and Efficient Cell Tagging with Block Copolymer-Based Contrast Agents for Sensitive MRI Detection in Vivo

“…This last factor is governed by size (6) and shape of the particles (14), but also by their surface functionalization (46), which largely regulates the interactions between the nanoparticles and the cells; nevertheless, special care must be taken to preserve the cellular functions of the labeled cells, since the migration of the administered cells remains interesting from a therapeutic point of view. MNP internalization is not a trivial issue, and therefore several strategies such as the use of transfection agents (1,4,23,28,42), cationic lipids (46), specific peptide functionalization (33), or electroporation (13,49) have been explored to increase the cellular uptake. In this work, we have refrained from using the above-mentioned methods and assessed the cell-labeling efficacies of P127- and T908-coated superparamagnetic nanoparticles (MNPs).…”