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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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BMSCs were analyzed using fluorescence-activated cell sorting and differentiation assays. Positive rates of expression of the third generation BMSCs of (A) CD29, (B) CD90 and (C) CD45 were 84.69, 90.28 and 0.72, respectively. BMSC, bone marrow stromal cell.
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f2-mmr-11-02-0954: BMSCs were analyzed using fluorescence-activated cell sorting and differentiation assays. Positive rates of expression of the third generation BMSCs of (A) CD29, (B) CD90 and (C) CD45 were 84.69, 90.28 and 0.72, respectively. BMSC, bone marrow stromal cell.

Mentions: The primary bone marrow stromal cells (BMSCs) were seeded onto petri dishes following spherical suspension in culture medium for 10 h for adherence. After 48 h of adherence, the cells had a stretched appearance with short spindles, triangular centered nuclei and marked refraction, rapidly demonstrating colony of amplification (Fig. 1). The expression of cell surface antigens was detected using flow cytometry. With proceeding incubation time, gradual necrosis was observed in the suspended hematopoietic cells; however, replacing the medium led to the growth of evenly distributed cells with a fusiform fibrotic shape. The expression of cell surface antigens was detected using flow cytometry. The flow cytometeric analysis of the BMSC surface antigens revealed CD29, CD45 and CD90 at 84.69, 0.72 and 90.28%, respectively (Fig. 2).


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)

BMSCs were analyzed using fluorescence-activated cell sorting and differentiation assays. Positive rates of expression of the third generation BMSCs of (A) CD29, (B) CD90 and (C) CD45 were 84.69, 90.28 and 0.72, respectively. BMSC, bone marrow stromal cell.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2-mmr-11-02-0954: BMSCs were analyzed using fluorescence-activated cell sorting and differentiation assays. Positive rates of expression of the third generation BMSCs of (A) CD29, (B) CD90 and (C) CD45 were 84.69, 90.28 and 0.72, respectively. BMSC, bone marrow stromal cell.
Mentions: The primary bone marrow stromal cells (BMSCs) were seeded onto petri dishes following spherical suspension in culture medium for 10 h for adherence. After 48 h of adherence, the cells had a stretched appearance with short spindles, triangular centered nuclei and marked refraction, rapidly demonstrating colony of amplification (Fig. 1). The expression of cell surface antigens was detected using flow cytometry. With proceeding incubation time, gradual necrosis was observed in the suspended hematopoietic cells; however, replacing the medium led to the growth of evenly distributed cells with a fusiform fibrotic shape. The expression of cell surface antigens was detected using flow cytometry. The flow cytometeric analysis of the BMSC surface antigens revealed CD29, CD45 and CD90 at 84.69, 0.72 and 90.28%, respectively (Fig. 2).

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