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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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Transmission electron microscopy (magnification, ×50,000) revealed labeled bone marrow stromal cells. Gadolinium diethylenetriamine pentaacetic acid particles were observed within the cytoplasm (arrow).
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f3-mmr-11-02-0954: Transmission electron microscopy (magnification, ×50,000) revealed labeled bone marrow stromal cells. Gadolinium diethylenetriamine pentaacetic acid particles were observed within the cytoplasm (arrow).

Mentions: Gd-DTPA is a small soluble molecule. Following mixing with effectene, numerous Gd-DTPA particles with a negative charge were observed, which were initially condensed together and were subsequently coated with cationic lipids and transferred into cells. The internalized Gd-DTPA particles were mainly aggregated together. Following labeling, the Gd-DTPA particles were clearly visible inside the cytoplasm and accentuated around the cellular apparatus. The high-density particles measured ~50 μm in diameter under the electron microscope (Fig. 3). Using atomic emission spectrophotometry, the effectene-mediated Gd-DTPA labeling efficiency was determined to be 86%.


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)

Transmission electron microscopy (magnification, ×50,000) revealed labeled bone marrow stromal cells. Gadolinium diethylenetriamine pentaacetic acid particles were observed within the cytoplasm (arrow).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f3-mmr-11-02-0954: Transmission electron microscopy (magnification, ×50,000) revealed labeled bone marrow stromal cells. Gadolinium diethylenetriamine pentaacetic acid particles were observed within the cytoplasm (arrow).
Mentions: Gd-DTPA is a small soluble molecule. Following mixing with effectene, numerous Gd-DTPA particles with a negative charge were observed, which were initially condensed together and were subsequently coated with cationic lipids and transferred into cells. The internalized Gd-DTPA particles were mainly aggregated together. Following labeling, the Gd-DTPA particles were clearly visible inside the cytoplasm and accentuated around the cellular apparatus. The high-density particles measured ~50 μm in diameter under the electron microscope (Fig. 3). Using atomic emission spectrophotometry, the effectene-mediated Gd-DTPA labeling efficiency was determined to be 86%.

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
Related in: MedlinePlus