Limits...
Dual-modal nanoprobes for imaging of mesenchymal stem cell transplant by MRI and fluorescence imaging.

Sung CK, Hong KA, Lin S, Lee Y, Cha J, Lee JK, Hong CP, Han BS, Jung SI, Kim SH, Yoon KS - Korean J Radiol (2009 Nov-Dec)

Bottom Line: The effective internalization of MNP@SiO(2)(RITC)-PEG into hMSCs was observed by confocal laser scanning fluorescence microscopy.MNP@SiO(2)(RITC)-PEG-labeled hMSCs were observed in vitro and in vivo with optical and MR imaging.MNP@SiO(2)(RITC)-PEG can be a useful contrast agent for stem cell imaging, which is suitable for a bimodal detection by MRI and optical imaging.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiology, Seoul Metropolitan Boramae Medical Center, Seoul National University College of Medicine, Seoul 156-707, Korea. sckmd@hanmail.net

ABSTRACT

Objective: To determine the feasibility of labeling human mesenchymal stem cells (hMSCs) with bifunctional nanoparticles and assessing their potential as imaging probes in the monitoring of hMSC transplantation.

Materials and methods: The T1 and T2 relaxivities of the nanoparticles (MNP@SiO(2)[RITC]-PEG) were measured at 1.5T and 3T magnetic resonance scanner. Using hMSCs and the nanoparticles, labeling efficiency, toxicity, and proliferation were assessed. Confocal laser scanning microscopy and transmission electron microscopy were used to specify the intracellular localization of the endocytosed iron nanoparticles. We also observed in vitro and in vivo visualization of the labeled hMSCs with a 3T MR scanner and optical imaging.

Results: MNP@SiO(2)(RITC)-PEG showed both superparamagnetic and fluorescent properties. The r(1) and r(2) relaxivity values of the MNP@SiO(2)(RITC)-PEG were 0.33 and 398 mM(-1) s(-1) at 1.5T, respectively, and 0.29 and 453 mM(-1) s(-1) at 3T, respectively. The effective internalization of MNP@SiO(2)(RITC)-PEG into hMSCs was observed by confocal laser scanning fluorescence microscopy. The transmission electron microscopy images showed that MNP@SiO(2)(RITC)-PEG was internalized into the cells and mainly resided in the cytoplasm. The viability and proliferation of MNP@SiO(2)(RITC)-PEG-labeled hMSCs were not significantly different from the control cells. MNP@SiO(2)(RITC)-PEG-labeled hMSCs were observed in vitro and in vivo with optical and MR imaging.

Conclusion: MNP@SiO(2)(RITC)-PEG can be a useful contrast agent for stem cell imaging, which is suitable for a bimodal detection by MRI and optical imaging.

Show MeSH

Related in: MedlinePlus

Cellular distribution of MNP@SiO2(RITC)-PEG in human mesenchymal stem cells. Confocal fluorescence images of human mesenchymal stem cells cultured with different concentrations of MNP@SiO2(RITC)-PEG (0, 0.5, 1, 2 and 4 mg/ml) for 2, 4, and 6 hours.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2770830&req=5

Figure 2: Cellular distribution of MNP@SiO2(RITC)-PEG in human mesenchymal stem cells. Confocal fluorescence images of human mesenchymal stem cells cultured with different concentrations of MNP@SiO2(RITC)-PEG (0, 0.5, 1, 2 and 4 mg/ml) for 2, 4, and 6 hours.

Mentions: MNP@SiO2(RITC)-PEG showed both superparamagnetic and fluorescent properties. The r1 and r2 relaxivity values of the MNP@SiO2(RITC)-PEG were 0.33 and 398 mM-1 s-1 at 1.5T, respectively, and 0.29 and 453 mM-1 s-1 at 3T, respectively (Fig. 1). The effective internalization of MNP@SiO2(RITC)-PEG into the hMSCs was observed by confocal laser scanning fluorescence microscopy as shown in Figure 2. In these fluorescence images, the hMSCs cultured with MNP@SiO2(RITC)-PEG took up a substantial amount of MNP@SiO2(RITC)-PEG as clearly identified by intracellular fluorescence signals, whereas those cultured without MNP@SiO2(RITC)-PEG were not detected by confocal LSM. There are no differences in the rate of uptake with increasing incubation concentrations of MNP@SiO2(RITC)-PEG. Confocal LSM showed that MNP@SiO2(RITC)-PEG particles resided in the cytoplasm and outside of the nucleus of a cell. These results confirmed that the MNP@SiO2(RITC)-PEG particles exhibited a strong role in targeting hMSCs and that the cellular uptake of MNP@SiO2(RITC)-PEG particles can be visualized by fluorescence imaging at the cellular level. We examined the localization and morphology of MNP@SiO2(RITC)-PEG particles by electron microscopy. The characteristic granular morphology of these MNP@SiO2(RITC)-PEG particles was identified only in MNP@SiO2(RITC)-PEG -labeled hMSCs and never in unlabeled controls. The TEM images showed that MNP@SiO2(RITC)-PEG were indeed internalized into the cells and mainly resided in the cytoplasm (Fig. 3).


Dual-modal nanoprobes for imaging of mesenchymal stem cell transplant by MRI and fluorescence imaging.

Sung CK, Hong KA, Lin S, Lee Y, Cha J, Lee JK, Hong CP, Han BS, Jung SI, Kim SH, Yoon KS - Korean J Radiol (2009 Nov-Dec)

Cellular distribution of MNP@SiO2(RITC)-PEG in human mesenchymal stem cells. Confocal fluorescence images of human mesenchymal stem cells cultured with different concentrations of MNP@SiO2(RITC)-PEG (0, 0.5, 1, 2 and 4 mg/ml) for 2, 4, and 6 hours.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2770830&req=5

Figure 2: Cellular distribution of MNP@SiO2(RITC)-PEG in human mesenchymal stem cells. Confocal fluorescence images of human mesenchymal stem cells cultured with different concentrations of MNP@SiO2(RITC)-PEG (0, 0.5, 1, 2 and 4 mg/ml) for 2, 4, and 6 hours.
Mentions: MNP@SiO2(RITC)-PEG showed both superparamagnetic and fluorescent properties. The r1 and r2 relaxivity values of the MNP@SiO2(RITC)-PEG were 0.33 and 398 mM-1 s-1 at 1.5T, respectively, and 0.29 and 453 mM-1 s-1 at 3T, respectively (Fig. 1). The effective internalization of MNP@SiO2(RITC)-PEG into the hMSCs was observed by confocal laser scanning fluorescence microscopy as shown in Figure 2. In these fluorescence images, the hMSCs cultured with MNP@SiO2(RITC)-PEG took up a substantial amount of MNP@SiO2(RITC)-PEG as clearly identified by intracellular fluorescence signals, whereas those cultured without MNP@SiO2(RITC)-PEG were not detected by confocal LSM. There are no differences in the rate of uptake with increasing incubation concentrations of MNP@SiO2(RITC)-PEG. Confocal LSM showed that MNP@SiO2(RITC)-PEG particles resided in the cytoplasm and outside of the nucleus of a cell. These results confirmed that the MNP@SiO2(RITC)-PEG particles exhibited a strong role in targeting hMSCs and that the cellular uptake of MNP@SiO2(RITC)-PEG particles can be visualized by fluorescence imaging at the cellular level. We examined the localization and morphology of MNP@SiO2(RITC)-PEG particles by electron microscopy. The characteristic granular morphology of these MNP@SiO2(RITC)-PEG particles was identified only in MNP@SiO2(RITC)-PEG -labeled hMSCs and never in unlabeled controls. The TEM images showed that MNP@SiO2(RITC)-PEG were indeed internalized into the cells and mainly resided in the cytoplasm (Fig. 3).

Bottom Line: The effective internalization of MNP@SiO(2)(RITC)-PEG into hMSCs was observed by confocal laser scanning fluorescence microscopy.MNP@SiO(2)(RITC)-PEG-labeled hMSCs were observed in vitro and in vivo with optical and MR imaging.MNP@SiO(2)(RITC)-PEG can be a useful contrast agent for stem cell imaging, which is suitable for a bimodal detection by MRI and optical imaging.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiology, Seoul Metropolitan Boramae Medical Center, Seoul National University College of Medicine, Seoul 156-707, Korea. sckmd@hanmail.net

ABSTRACT

Objective: To determine the feasibility of labeling human mesenchymal stem cells (hMSCs) with bifunctional nanoparticles and assessing their potential as imaging probes in the monitoring of hMSC transplantation.

Materials and methods: The T1 and T2 relaxivities of the nanoparticles (MNP@SiO(2)[RITC]-PEG) were measured at 1.5T and 3T magnetic resonance scanner. Using hMSCs and the nanoparticles, labeling efficiency, toxicity, and proliferation were assessed. Confocal laser scanning microscopy and transmission electron microscopy were used to specify the intracellular localization of the endocytosed iron nanoparticles. We also observed in vitro and in vivo visualization of the labeled hMSCs with a 3T MR scanner and optical imaging.

Results: MNP@SiO(2)(RITC)-PEG showed both superparamagnetic and fluorescent properties. The r(1) and r(2) relaxivity values of the MNP@SiO(2)(RITC)-PEG were 0.33 and 398 mM(-1) s(-1) at 1.5T, respectively, and 0.29 and 453 mM(-1) s(-1) at 3T, respectively. The effective internalization of MNP@SiO(2)(RITC)-PEG into hMSCs was observed by confocal laser scanning fluorescence microscopy. The transmission electron microscopy images showed that MNP@SiO(2)(RITC)-PEG was internalized into the cells and mainly resided in the cytoplasm. The viability and proliferation of MNP@SiO(2)(RITC)-PEG-labeled hMSCs were not significantly different from the control cells. MNP@SiO(2)(RITC)-PEG-labeled hMSCs were observed in vitro and in vivo with optical and MR imaging.

Conclusion: MNP@SiO(2)(RITC)-PEG can be a useful contrast agent for stem cell imaging, which is suitable for a bimodal detection by MRI and optical imaging.

Show MeSH
Related in: MedlinePlus