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Tracking of mesenchymal stem cells labeled with gadolinium diethylenetriamine pentaacetic acid by 7T magnetic resonance imaging in a model of cerebral ischemia.

Geng K, Yang ZX, Huang D, Yi M, Jia Y, Yan G, Cheng X, Wu R - Mol Med Rep (2014)

Bottom Line: The non‑liposomal lipid transfection reagent effectene was then used to induce the intracellular uptake of Gd‑DTPA.The T1‑weighted imaging of the labeled cells revealed a significantly higher signal intensity compared with that of the unlabeled cells (P<0.05) and the T1 values were significantly lower.The function of the labeled MSCs demonstrated no change following Gd‑DTPA labeling, with no evident adverse effect on cell viability or proliferation.

View Article: PubMed Central - PubMed

Affiliation: The Chinese People's Liberation Army 59 Hospital, Yunnan, Kaiyuan, Yunnan 661699, P.R. China.

ABSTRACT
Progress in the development of stem cell and gene therapy requires repeatable and non‑invasive techniques to monitor the survival and integration of stem cells in vivo with a high temporal and spatial resolution. The purpose of the present study was to examine the feasibility of using the standard contrast agent gadolinium diethylenetriamine pentaacetic acid (Gd‑DTPA) to label rat mesenchymal stem cells (MSCs) for stem cell tracking. MSCs, obtained from the bilateral femora of rats, were cultured and propagated. The non‑liposomal lipid transfection reagent effectene was then used to induce the intracellular uptake of Gd‑DTPA. Electron microscopy was used to detect the distribution of Gd‑DTPA particles in the MSCs. The labeling efficiency of the Gd‑DTPA particles in the MSCs was determined using spectrophotometry, and MTT and trypan blue exclusion assays were used to evaluate the viability and proliferation of the labeled MSCs. T1‑weighted magnetic resonance imaging (MRI) was used to observe the labeled cells in vitro and in the rat brain. Gd‑DTPA particles were detected inside the MSCs using transmission electron microscopy and a high labeling efficiency was observed. No difference was observed in cell viability or proliferation between the labeled and unlabeled MSCs (P>0.05). In the in vitro T1‑weighted MRI and in the rat brain, a high signal intensity was observed in the labeled MSCs. The T1‑weighted imaging of the labeled cells revealed a significantly higher signal intensity compared with that of the unlabeled cells (P<0.05) and the T1 values were significantly lower. The function of the labeled MSCs demonstrated no change following Gd‑DTPA labeling, with no evident adverse effect on cell viability or proliferation. Therefore, a change in MR signal intensity was detected in vitro and in vivo, suggesting Gd‑DTPA can be used to label MSCs for MRI tracking.

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Imaging of cells using magentic resonance in vitro. The T1-weighted spin echo image revealed that the signal intensity of (A) the group containing Gd-DTPA, effectene and bone marrow stromal cells was markedly higher compared with that of (B) the control groups or groups treated with (C) Gd-DTPA or (D) effectene only. Gd-DTPA, gadolinium diethylenetriamine pentaacetic acid.
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f5-mmr-11-02-0954: Imaging of cells using magentic resonance in vitro. The T1-weighted spin echo image revealed that the signal intensity of (A) the group containing Gd-DTPA, effectene and bone marrow stromal cells was markedly higher compared with that of (B) the control groups or groups treated with (C) Gd-DTPA or (D) effectene only. Gd-DTPA, gadolinium diethylenetriamine pentaacetic acid.

Mentions: As shown in Fig. 5, the labeled cells (Fig. 5A) demonstrated markedly higher signal intensity in T1-weighted imaging compared with the unlabeled cells (Fig. 5B) and the cells incubated with either Gd-DTPA (Fig. 5C) or effectene alone (Fig. 5D). This signal feature was consistent with the presence of Gd-DTPA particles inside the cytoplasm. Subsequently, the increased signal intensity of T1-weighted imaging of the labeled cells was evaluated and the T1 values of the labeled cells were further measured. In T1-weighted imaging, the signal intensities of the labeled cells, the cells with Gd-DTPA, the cells with effectene and the unlabeled cells were 1067±34, 635±16, 603±19 and 598±25, respectively. The signal intensity of the labeled cells was increased significantly compared with that of the negative control cells (P<0.001) and the signal intensity of the labeled cells was 1.48 times higher than that of the unlabeled cells. The T1 values of the labeled cells, the cells with Gd-DTPA, the cells with effectene and the unlabeled cells were 886±269, 2,086±58, 2,182±280 and 2,185±200 ms, respectively. The T1 value of the labeled cells was decreased significantly compared with those of the negative controls and the unlabeled cells (P<0.001). The T1 values of the labeled cells were 2.46 times lower than those of the unlabeled cells. In terms of cell proliferation, the T1-weighted image signal intensity of the BMSCs reduced gradually and, at the fifth generation, the signal intensity was similar to that observed in the unlabeled cells (Fig. 6).


Tracking of mesenchymal stem cells labeled with gadolinium diethylenetriamine pentaacetic acid by 7T magnetic resonance imaging in a model of cerebral ischemia.

Geng K, Yang ZX, Huang D, Yi M, Jia Y, Yan G, Cheng X, Wu R - Mol Med Rep (2014)

Imaging of cells using magentic resonance in vitro. The T1-weighted spin echo image revealed that the signal intensity of (A) the group containing Gd-DTPA, effectene and bone marrow stromal cells was markedly higher compared with that of (B) the control groups or groups treated with (C) Gd-DTPA or (D) effectene only. Gd-DTPA, gadolinium diethylenetriamine pentaacetic acid.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4262487&req=5

f5-mmr-11-02-0954: Imaging of cells using magentic resonance in vitro. The T1-weighted spin echo image revealed that the signal intensity of (A) the group containing Gd-DTPA, effectene and bone marrow stromal cells was markedly higher compared with that of (B) the control groups or groups treated with (C) Gd-DTPA or (D) effectene only. Gd-DTPA, gadolinium diethylenetriamine pentaacetic acid.
Mentions: As shown in Fig. 5, the labeled cells (Fig. 5A) demonstrated markedly higher signal intensity in T1-weighted imaging compared with the unlabeled cells (Fig. 5B) and the cells incubated with either Gd-DTPA (Fig. 5C) or effectene alone (Fig. 5D). This signal feature was consistent with the presence of Gd-DTPA particles inside the cytoplasm. Subsequently, the increased signal intensity of T1-weighted imaging of the labeled cells was evaluated and the T1 values of the labeled cells were further measured. In T1-weighted imaging, the signal intensities of the labeled cells, the cells with Gd-DTPA, the cells with effectene and the unlabeled cells were 1067±34, 635±16, 603±19 and 598±25, respectively. The signal intensity of the labeled cells was increased significantly compared with that of the negative control cells (P<0.001) and the signal intensity of the labeled cells was 1.48 times higher than that of the unlabeled cells. The T1 values of the labeled cells, the cells with Gd-DTPA, the cells with effectene and the unlabeled cells were 886±269, 2,086±58, 2,182±280 and 2,185±200 ms, respectively. The T1 value of the labeled cells was decreased significantly compared with those of the negative controls and the unlabeled cells (P<0.001). The T1 values of the labeled cells were 2.46 times lower than those of the unlabeled cells. In terms of cell proliferation, the T1-weighted image signal intensity of the BMSCs reduced gradually and, at the fifth generation, the signal intensity was similar to that observed in the unlabeled cells (Fig. 6).

Bottom Line: The non‑liposomal lipid transfection reagent effectene was then used to induce the intracellular uptake of Gd‑DTPA.The T1‑weighted imaging of the labeled cells revealed a significantly higher signal intensity compared with that of the unlabeled cells (P<0.05) and the T1 values were significantly lower.The function of the labeled MSCs demonstrated no change following Gd‑DTPA labeling, with no evident adverse effect on cell viability or proliferation.

View Article: PubMed Central - PubMed

Affiliation: The Chinese People's Liberation Army 59 Hospital, Yunnan, Kaiyuan, Yunnan 661699, P.R. China.

ABSTRACT
Progress in the development of stem cell and gene therapy requires repeatable and non‑invasive techniques to monitor the survival and integration of stem cells in vivo with a high temporal and spatial resolution. The purpose of the present study was to examine the feasibility of using the standard contrast agent gadolinium diethylenetriamine pentaacetic acid (Gd‑DTPA) to label rat mesenchymal stem cells (MSCs) for stem cell tracking. MSCs, obtained from the bilateral femora of rats, were cultured and propagated. The non‑liposomal lipid transfection reagent effectene was then used to induce the intracellular uptake of Gd‑DTPA. Electron microscopy was used to detect the distribution of Gd‑DTPA particles in the MSCs. The labeling efficiency of the Gd‑DTPA particles in the MSCs was determined using spectrophotometry, and MTT and trypan blue exclusion assays were used to evaluate the viability and proliferation of the labeled MSCs. T1‑weighted magnetic resonance imaging (MRI) was used to observe the labeled cells in vitro and in the rat brain. Gd‑DTPA particles were detected inside the MSCs using transmission electron microscopy and a high labeling efficiency was observed. No difference was observed in cell viability or proliferation between the labeled and unlabeled MSCs (P>0.05). In the in vitro T1‑weighted MRI and in the rat brain, a high signal intensity was observed in the labeled MSCs. The T1‑weighted imaging of the labeled cells revealed a significantly higher signal intensity compared with that of the unlabeled cells (P<0.05) and the T1 values were significantly lower. The function of the labeled MSCs demonstrated no change following Gd‑DTPA labeling, with no evident adverse effect on cell viability or proliferation. Therefore, a change in MR signal intensity was detected in vitro and in vivo, suggesting Gd‑DTPA can be used to label MSCs for MRI tracking.

Show MeSH