Abstract
Background aims. Mesenchymal stromal cells (MSC) are the focus of research in regenerative medicine aiming at the regulatory approval of these cells for specific indications. To cope with the regulatory requirements for somatic cell therapy, novel approaches that do not interfere with the natural behavior of the cells are necessary. In this context in vivo magnetic resonance imaging (MRI) of labeled MSC could be an appropriate tool. Cell labeling for MRI with a variety of different iron oxide preparations is frequently published. However, most publications lack a comprehensive assessment of the noninterference of the contrast agent with the functionality of the labeled MSC, which is a prerequisite for the validity of cell-tracking via MRI. Methods.We studied the effects of iron oxide-poly(L-lactide) nanoparticles in MSC with flow cytom-etry, transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), Prussian blue staining, CyQuant® proliferation testing, colony-forming unit-fibroblast (CFU-F) assays, flow chamber adhesion testing, immuno-logic tests and differentiation tests. Furthermore iron-labeled MSC were studied by MRI in agarose phantoms and Wistar rats. Results. It could be demonstrated that MSC show rapid uptake of nanoparticles and long-lasting intracellular persistence in the endosomal compartment. Labeling of the MSC with these particles has no influence on viability, differentiation, clonogenicity, proliferation, adhesion, phenotype and immunosuppressive properties. They show excellent MRI properties in agarose phantoms and after subcutaneous implantation in rats over several weeks. Conclusions. These particles qualify for studying MSC homing and trafficking via MRI.
Highlights
Mesenchymal stromal/stem cells (MSC) are the focus of interest in regenerative medicine, nurtured by their proven ability to differentiate into different cell types deriving from the mesoderm and their abundant availability because they are easy to culture and expand in vitro [1,2]
In the study presented here, we investigated MSC labeling with iron oxide–poly(L-lactide) (PLLA) nanoparticles synthesized via the mini-emulsion process [35,36]
Persistence and toxicity measurements, MSC were incubated for 24 h with nanoparticles and evaluated by transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), fluorescentactivated cell sorting (FACS) and Prussian blue staining
Summary
Mesenchymal stromal/stem cells (MSC) are the focus of interest in regenerative medicine, nurtured by their proven ability to differentiate into different cell types deriving from the mesoderm and their abundant availability because they are easy to culture and expand in vitro [1,2]. In the study presented here, we investigated MSC labeling with iron oxide–poly(L-lactide) (PLLA) nanoparticles synthesized via the mini-emulsion process [35,36]. To cope with the regulatory requirements for somatic cell therapy, novel approaches that do not interfere with the natural behavior of the cells are necessary In this context in vivo magnetic resonance imaging (MRI) of labeled MSC could be an appropriate tool. Labeling of the MSC with these particles has no influence on viability, differentiation, clonogenicity, proliferation, adhesion, phenotype and immunosuppressive properties. They show excellent MRI properties in agarose phantoms and after subcutaneous implantation in rats over several weeks. These particles qualify for studying MSC homing and trafficking via MRI
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