Korean J Radiol.  2007 Oct;8(5):365-371. 10.3348/kjr.2007.8.5.365.

Labeling Efficacy of Superparamagnetic Iron Oxide Nanoparticles to Human Neural Stem Cells: Comparison of Ferumoxides, Monocrystalline Iron Oxide, Cross-linked Iron Oxide (CLIO)-NH2 and tat-CLIO

• Song M
• Moon WK
• Kim Y
• Lim D
• Song IC
• Yoon BW

• KMID: 1734284
• DOI: http://doi.org/10.3348/kjr.2007.8.5.365

Abstract

OBJECTIVE
We wanted to compare the human neural stem cell (hNSC) labeling efficacy of different superparamagnetic iron oxide nanoparticles (SPIONs), namely, ferumoxides, monocrystalline iron oxide (MION), cross-linked iron oxide (CLIO)-NH2 and tat-CLIO. MATERIALS AND METHODS: The hNSCs (5x105 HB1F3 cells/ml) were incubated for 24 hr in cell culture media that contained 25 microgram/ml of ferumoxides, MION or CLIO-NH2, and with or without poly-L-lysine (PLL) and tat-CLIO. The cellular iron uptake was analyzed qualitatively with using a light microscope and this was quantified via atomic absorption spectrophotometry. The visibility of the labeled cells was assessed with MR imaging. RESULTS: The incorporation of SPIONs into the hNSCs did not affect the cellular proliferations and viabilities. The hNSCs labeled with tat-CLIO showed the longest retention, up to 72 hr, and they contained 2.15+/-0.3 pg iron/cell, which are 59 fold, 430 fold and six fold more incorporated iron than that of the hNSCs labeled with ferumoxides, MION or CLIO-NH2, respectively. However, when PLL was added, the incorporation of ferumoxides, MION or CLIO-NH2 into the hNSCs was comparable to that of tat-CLIO. CONCLUSION: For MR imaging, hNSCs can be efficiently labeled with tat-CLIO alone or with a combination of ferumoxides, MION, CLIO-NH2 and the transfection agent PLL.

Keyword

Human neural stem cell; Iron oxide nanoparticles; Magnetic resonance (MR)

MeSH Terms

Cells, Cultured
Contrast Media/chemical synthesis/pharmacokinetics
Cross-Linking Reagents/chemistry
Ferric Compounds/chemistry/*pharmacokinetics
Ferrosoferric Oxide/chemical synthesis/pharmacokinetics
Gene Products, tat/chemistry
Humans
Iron/*pharmacokinetics
Magnetic Resonance Imaging/methods
Nanoparticles
Neural Tube
Oxides/*pharmacokinetics
Phantoms, Imaging
Polylysine/pharmacokinetics
Spectrophotometry, Atomic
Staining and Labeling/*methods
Stem Cells/cytology/*drug effects/metabolism
Time Factors
Transfection

Figure

• 

  Fig. 1 Photomicrographs of the hNSCs treated for 24 hr with ferumoxides (A), MION-47 (B), CLIO-NH2 (C) or tat-CLIO (D) at 25 µg/ml. The intracellular uptake of iron oxide nanoparticles (arrows) is seen in cells exposed to ferumoxides (A), CLIO-NH2 (C) or tat-CLIO (D). However, no intracellular uptake of iron oxide was found for the cells incubated with MION-47 (B). (Prussian blue stain, objective magnification: × 40)

• 

  Fig. 2 Retention of SPIONs in hNSCs. Iron oxide nanoparticles (arrows) are seen within the ferumoxides labeled cells and the CLIO-NH2 labeled cells for up to 24 and 48 hr, respectively and inside the tat-CLIO labeled cells for up to 72 hr. (Prussian blue stain, objective magnification: × 40)

• 

  Fig. 3 Photomicrographs of hNSCs treated for 24 hr with three different SPIONs (ferumoxides, MION-47 or CLIO-NH2) at 25 µg/ml in the presence of different doses of PLL. As the PLL concentration increased in the media from 0.25 µg/ml to 2 µg/ml, more iron oxide nanoparticles are seen inside the labeled cells. (Prussian blue stain, objective magnification: × 40)

• 

  Fig. 4 Quantitative iron determination by an atomic absorption spectrophotometer. The graph shows the highest iron incorporation in the tat-CLIO labeled cells (2.15 ± 0.3 pg/cell) and the lowest in the MION-47 labeled cells (0.005 pg/cell) in absence of PLL (black bars). With PLL (white bars), the iron content in the ferumoxides labeled, MION-47 labeled or CLIO-NH2 labeled cells increased to 1.08 ± 0.07 pg/cell, 1.01 ± 0.02 pg/cell and 2.24 ± 0.17 pg/cell, respectively, which are 27 fold, 202 fold and 7-fold greater uptakes, respectively, compared with the cells without using PLL. The cells labeled with CLIO-NH2 in the presence of PLL show a similar intracellular iron content as the tat-CLIO-labeled cells (p > 0.1). Note.-w/o = without, w = with, ns = statistically non-significant

• 

  Fig. 5 Gradient-echo T2-weighted MR images of the phantoms with different cell numbers (i.e., 312, 625, 1,250, 2,500, 5,000 and 10,000 cells/µl). MR scans were performed at different time points with (A) the control, the MION-47 treated cells and the MION-47+PLL treated cells, (B) the ferumoxides treated cells and the ferumoxides with PLL treated cells, and (C) the CLIO-NH2 treated cells, the CLIO-NH2 with PLL treated cells and the tat-CLIO treated cells. In the absence of PLL, the phantoms with more than 1,250 cells/µl of the tat-CLIO-labeled cells show a decreased signal on the T2-weighted MR images. For the ferumoxides treated cells and the CLIO-NH2 treated cells, the phantoms with more than 2,500 cells/µl show a visibly decreased signal without PLL. With PLL, a decreased signal is found in the phantoms with the ferumoxides treated cells, in the phantoms with the MION-47 treated cells and in the phantoms with the CLIO-NH2 treated cells, with 625-1,250 cells/µl respectively. 'Control' represents the MR image of the phantom with the non-labeled control cells.

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