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


CONCLUSIONS
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.
Instituto do Cérebro, Hospital Israelita Albert Einstein, São Paulo, SP, Brazil.
Tatiana Taís
TT
Sibov
Liza Aya Mabuchi
LA
Miyaki
Javier Bustamante
JB
Mamani
Luciana Cavalheiro
LC
Marti
Luiz Roberto
LR
Sardinha
Lorena Favaro
LF
Pavon
Daniela Mara de
DM
Oliveira
Walter Humberto
WH
Cardenas
Lionel Fernel
LF
Gamarra
D049109:Cell Proliferation; D058948:Cell Tracking / Q000379:methods*; D003911:Dextrans / Q000737:chemistry*; D005290:Ferric Compounds; D005434:Flow Cytometry; D006801:Humans; D058185:Magnetite Nanoparticles; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D011107:Polylysine / Q000737:chemistry*; D013194:Staining and Labeling; D014470:Umbilical Cord / Q000166:cytology*
D003911:Dextrans; D005290:Ferric Compounds; D058185:Magnetite Nanoparticles; C000499:ferric oxide; D011107:Polylysine