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Labeling of mesenchymal stem cells for MRI with single-cell sensitivity.

Ariza de Schellenberger A, Kratz H, Farr TD, Löwa N, Hauptmann R, Wagner S, Taupitz M, Schnorr J, Schellenberger EA - Int J Nanomedicine (2016)

Bottom Line: Attempts should be made to achieve better cell separation for homogeneous NP loading and to thus improve NP-uptake-dependent biocompatibility studies and cell detection by MRI and future MPI.Additionally, using a 7 T MR imager equipped with a cryocoil resulted in approximately two times higher detection.In conclusion, we established labeling conditions for new high-relaxivity MCP, VSOP, and Resovist(®) for improved MRI of MSC with single-cell sensitivity.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiology, Center for Stroke Research Berlin, Charité - Universitätsmedizin Berlin, Berlin, Germany.

ABSTRACT
Sensitive cell detection by magnetic resonance imaging (MRI) is an important tool for the development of cell therapies. However, clinically approved contrast agents that allow single-cell detection are currently not available. Therefore, we compared very small iron oxide nanoparticles (VSOP) and new multicore carboxymethyl dextran-coated iron oxide nanoparticles (multicore particles, MCP) designed by our department for magnetic particle imaging (MPI) with discontinued Resovist(®) regarding their suitability for detection of single mesenchymal stem cells (MSC) by MRI. We achieved an average intracellular nanoparticle (NP) load of >10 pg Fe per cell without the use of transfection agents. NP loading did not lead to significantly different results in proliferation, colony formation, and multilineage in vitro differentiation assays in comparison to controls. MRI allowed single-cell detection using VSOP, MCP, and Resovist(®) in conjunction with high-resolution T2*-weighted imaging at 7 T with postprocessing of phase images in agarose cell phantoms and in vivo after delivery of 2,000 NP-labeled MSC into mouse brains via the left carotid artery. With optimized labeling conditions, a detection rate of ~45% was achieved; however, the experiments were limited by nonhomogeneous NP loading of the MSC population. Attempts should be made to achieve better cell separation for homogeneous NP loading and to thus improve NP-uptake-dependent biocompatibility studies and cell detection by MRI and future MPI. Additionally, using a 7 T MR imager equipped with a cryocoil resulted in approximately two times higher detection. In conclusion, we established labeling conditions for new high-relaxivity MCP, VSOP, and Resovist(®) for improved MRI of MSC with single-cell sensitivity.

No MeSH data available.


Related in: MedlinePlus

In vitro assays for nanoparticle cell biocompatibility.Notes: Cells labeled with Resovist®, MCP, or VSOP show similar population doubling time as unlabeled cells (empty) over 10 days (Days 2, 4, 6, and 10 are shown) (mean; bars, ± SD; n=3) (A). Cells maintained both their mesenchymal stem cell (MSC) character and their self-renewal capacity, showing similar number of colony-forming units (CFU) for labeled and unlabeled cells (B). Multipotent differentiation potential (adipogenesis, chondrogenesis, and osteogenesis) of labeled MSC was similar to unlabeled cells as shown by Oil Red, Alcian Blue, and von Kossa stains (C). All scale bars represent 100 µm.Abbreviations: MCP, multicore carboxy-methyl-dextran-coated iron oxide nanoparticle; VSOP, very small iron oxide nanoparticle.
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f3-ijn-11-1517: In vitro assays for nanoparticle cell biocompatibility.Notes: Cells labeled with Resovist®, MCP, or VSOP show similar population doubling time as unlabeled cells (empty) over 10 days (Days 2, 4, 6, and 10 are shown) (mean; bars, ± SD; n=3) (A). Cells maintained both their mesenchymal stem cell (MSC) character and their self-renewal capacity, showing similar number of colony-forming units (CFU) for labeled and unlabeled cells (B). Multipotent differentiation potential (adipogenesis, chondrogenesis, and osteogenesis) of labeled MSC was similar to unlabeled cells as shown by Oil Red, Alcian Blue, and von Kossa stains (C). All scale bars represent 100 µm.Abbreviations: MCP, multicore carboxy-methyl-dextran-coated iron oxide nanoparticle; VSOP, very small iron oxide nanoparticle.

Mentions: We chose to continue further experiments with highly labeled MSC without PS to test the biocompatibility of these labeling protocols in comparison with unlabeled cells. Assays for PDT, in vitro cell differentiation, CFU-F (Figure 3), and MSC marker expression (Figure S1) were performed. Overall, PDT assessed for 10 days after NP uptake was similar for labeled and unlabeled cells (Figure 3A). Labeled MSC showed a similar CFU capacity compared to unlabeled cells (Figure 3B). Furthermore, characteristic pluripotent features of MSC were not modified by VSOP or MCP labeling. Therefore, adipogenic, osteogenic, and chondrogenic differentiation could be similarly induced in labeled and unlabeled cells after in vitro stimulation (Figure 3C).


Labeling of mesenchymal stem cells for MRI with single-cell sensitivity.

Ariza de Schellenberger A, Kratz H, Farr TD, Löwa N, Hauptmann R, Wagner S, Taupitz M, Schnorr J, Schellenberger EA - Int J Nanomedicine (2016)

In vitro assays for nanoparticle cell biocompatibility.Notes: Cells labeled with Resovist®, MCP, or VSOP show similar population doubling time as unlabeled cells (empty) over 10 days (Days 2, 4, 6, and 10 are shown) (mean; bars, ± SD; n=3) (A). Cells maintained both their mesenchymal stem cell (MSC) character and their self-renewal capacity, showing similar number of colony-forming units (CFU) for labeled and unlabeled cells (B). Multipotent differentiation potential (adipogenesis, chondrogenesis, and osteogenesis) of labeled MSC was similar to unlabeled cells as shown by Oil Red, Alcian Blue, and von Kossa stains (C). All scale bars represent 100 µm.Abbreviations: MCP, multicore carboxy-methyl-dextran-coated iron oxide nanoparticle; VSOP, very small iron oxide nanoparticle.
© Copyright Policy
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4835118&req=5

f3-ijn-11-1517: In vitro assays for nanoparticle cell biocompatibility.Notes: Cells labeled with Resovist®, MCP, or VSOP show similar population doubling time as unlabeled cells (empty) over 10 days (Days 2, 4, 6, and 10 are shown) (mean; bars, ± SD; n=3) (A). Cells maintained both their mesenchymal stem cell (MSC) character and their self-renewal capacity, showing similar number of colony-forming units (CFU) for labeled and unlabeled cells (B). Multipotent differentiation potential (adipogenesis, chondrogenesis, and osteogenesis) of labeled MSC was similar to unlabeled cells as shown by Oil Red, Alcian Blue, and von Kossa stains (C). All scale bars represent 100 µm.Abbreviations: MCP, multicore carboxy-methyl-dextran-coated iron oxide nanoparticle; VSOP, very small iron oxide nanoparticle.
Mentions: We chose to continue further experiments with highly labeled MSC without PS to test the biocompatibility of these labeling protocols in comparison with unlabeled cells. Assays for PDT, in vitro cell differentiation, CFU-F (Figure 3), and MSC marker expression (Figure S1) were performed. Overall, PDT assessed for 10 days after NP uptake was similar for labeled and unlabeled cells (Figure 3A). Labeled MSC showed a similar CFU capacity compared to unlabeled cells (Figure 3B). Furthermore, characteristic pluripotent features of MSC were not modified by VSOP or MCP labeling. Therefore, adipogenic, osteogenic, and chondrogenic differentiation could be similarly induced in labeled and unlabeled cells after in vitro stimulation (Figure 3C).

Bottom Line: Attempts should be made to achieve better cell separation for homogeneous NP loading and to thus improve NP-uptake-dependent biocompatibility studies and cell detection by MRI and future MPI.Additionally, using a 7 T MR imager equipped with a cryocoil resulted in approximately two times higher detection.In conclusion, we established labeling conditions for new high-relaxivity MCP, VSOP, and Resovist(®) for improved MRI of MSC with single-cell sensitivity.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiology, Center for Stroke Research Berlin, Charité - Universitätsmedizin Berlin, Berlin, Germany.

ABSTRACT
Sensitive cell detection by magnetic resonance imaging (MRI) is an important tool for the development of cell therapies. However, clinically approved contrast agents that allow single-cell detection are currently not available. Therefore, we compared very small iron oxide nanoparticles (VSOP) and new multicore carboxymethyl dextran-coated iron oxide nanoparticles (multicore particles, MCP) designed by our department for magnetic particle imaging (MPI) with discontinued Resovist(®) regarding their suitability for detection of single mesenchymal stem cells (MSC) by MRI. We achieved an average intracellular nanoparticle (NP) load of >10 pg Fe per cell without the use of transfection agents. NP loading did not lead to significantly different results in proliferation, colony formation, and multilineage in vitro differentiation assays in comparison to controls. MRI allowed single-cell detection using VSOP, MCP, and Resovist(®) in conjunction with high-resolution T2*-weighted imaging at 7 T with postprocessing of phase images in agarose cell phantoms and in vivo after delivery of 2,000 NP-labeled MSC into mouse brains via the left carotid artery. With optimized labeling conditions, a detection rate of ~45% was achieved; however, the experiments were limited by nonhomogeneous NP loading of the MSC population. Attempts should be made to achieve better cell separation for homogeneous NP loading and to thus improve NP-uptake-dependent biocompatibility studies and cell detection by MRI and future MPI. Additionally, using a 7 T MR imager equipped with a cryocoil resulted in approximately two times higher detection. In conclusion, we established labeling conditions for new high-relaxivity MCP, VSOP, and Resovist(®) for improved MRI of MSC with single-cell sensitivity.

No MeSH data available.


Related in: MedlinePlus