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Simple SPION incubation as an efficient intracellular labeling method for tracking neural progenitor cells using MRI.

Chen CC, Ku MC, D M J, Lai JS, Hueng DY, Chang C - PLoS ONE (2013)

Bottom Line: However, compelling evidence also shows that simple SPION incubation is not invariably ineffective.The labeling efficiency can be improved by prolonged incubation and elevated iron doses.The results showed that, following 48 hours of incubation at 75 µg/ml, nearly all NPCs exhibited visible SPION intake.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biomedical Sciences, Academic Sinica, Taipei, Taiwan.

ABSTRACT
Cellular magnetic resonance imaging (MRI) has been well-established for tracking neural progenitor cells (NPC). Superparamagnetic iron oxide nanoparticles (SPIONs) approved for clinical application are the most common agents used for labeling. Conventionally, transfection agents (TAs) were added with SPIONs to facilitate cell labeling because SPIONs in the native unmodified form were deemed inefficient for intracellular labeling. However, compelling evidence also shows that simple SPION incubation is not invariably ineffective. The labeling efficiency can be improved by prolonged incubation and elevated iron doses. The goal of the present study was to establish simple SPION incubation as an efficient intracellular labeling method. To this end, NPCs derived from the neonatal subventricular zone were incubated with SPIONs (Feridex®) and then evaluated in vitro with regard to the labeling efficiency and biological functions. The results showed that, following 48 hours of incubation at 75 µg/ml, nearly all NPCs exhibited visible SPION intake. Evidence from light microscopy, electron microscopy, chemical analysis, and magnetic resonance imaging confirmed the effectiveness of the labeling. Additionally, biological assays showed that the labeled NPCs exhibited unaffected viability, oxidative stress, apoptosis and differentiation. In the demonstrated in vivo cellular MRI experiment, the hypointensities representing the SPION labeled NPCs remained observable throughout the entire tracking period. The findings indicate that simple SPION incubation without the addition of TAs is an efficient intracellular magnetic labeling method. This simple approach may be considered as an alternative approach to the mainstream labeling method that involves the use of TAs.

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A demonstration in vivo cellular MRI using the simple labeling method.A rat with glioma was transplanted with SPION-labeled NPCs into the lateral ventricle, and T2WIs were used to track the migration and distribution of the transplanted NPCs. (A) T2WI acquired at baseline, on day 1, 7, and 14 after transplantation. Hypointensities were readily visible one day later, and persisted throughout the tracking duration. (B) Signals at the tumor or injection site in relation to a control area (the cortex) were plotted against time. (C) PB staining confirmed the relocation of SPION-labeled cells following transplantation. Histology was performed on brain tissues obtained on day 14 after transplantation. The white arrow indicates the location of the tumor whereas the red arrows indicate the hypointensity arising from the labeled NPCs. For tumors without injected NPCs, the signal patterns were different. Untreated ENU tumors exhibited only mild hypointensities likely associated with hemorrhage on T2WI and T2*WI.
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pone-0056125-g007: A demonstration in vivo cellular MRI using the simple labeling method.A rat with glioma was transplanted with SPION-labeled NPCs into the lateral ventricle, and T2WIs were used to track the migration and distribution of the transplanted NPCs. (A) T2WI acquired at baseline, on day 1, 7, and 14 after transplantation. Hypointensities were readily visible one day later, and persisted throughout the tracking duration. (B) Signals at the tumor or injection site in relation to a control area (the cortex) were plotted against time. (C) PB staining confirmed the relocation of SPION-labeled cells following transplantation. Histology was performed on brain tissues obtained on day 14 after transplantation. The white arrow indicates the location of the tumor whereas the red arrows indicate the hypointensity arising from the labeled NPCs. For tumors without injected NPCs, the signal patterns were different. Untreated ENU tumors exhibited only mild hypointensities likely associated with hemorrhage on T2WI and T2*WI.

Mentions: The T2*WI and T2WI at baseline and at 1, 7, and 14 days after transplantation of SPION-labeled NPCs into the lateral ventricle of the rat with glioma are shown in Fig. 7A. The tumor was identified as a hyperintensity at baseline. One day following transplantation, the signal at the tumor site became hypointense and persisted throughout the tracking duration. Quantification of the signal at the tumor or injection site over time is shown in Fig. 7B. The tumor exhibited increasing hypointensity and the injection site remained the same hypointense. PB staining performed on the brain tissues obtained 14 days after transplantation indicated that iron was present in the tumor site, which should be responsible for the signal changes on T2*WI and T2WI (Fig. 7C). For tumors without injected NPCs, the signal patterns were different. Untreated ENU tumors exhibited only mild hypointensities associated with hemorrhage on T2WI and T2*WI (data not shown).


Simple SPION incubation as an efficient intracellular labeling method for tracking neural progenitor cells using MRI.

Chen CC, Ku MC, D M J, Lai JS, Hueng DY, Chang C - PLoS ONE (2013)

A demonstration in vivo cellular MRI using the simple labeling method.A rat with glioma was transplanted with SPION-labeled NPCs into the lateral ventricle, and T2WIs were used to track the migration and distribution of the transplanted NPCs. (A) T2WI acquired at baseline, on day 1, 7, and 14 after transplantation. Hypointensities were readily visible one day later, and persisted throughout the tracking duration. (B) Signals at the tumor or injection site in relation to a control area (the cortex) were plotted against time. (C) PB staining confirmed the relocation of SPION-labeled cells following transplantation. Histology was performed on brain tissues obtained on day 14 after transplantation. The white arrow indicates the location of the tumor whereas the red arrows indicate the hypointensity arising from the labeled NPCs. For tumors without injected NPCs, the signal patterns were different. Untreated ENU tumors exhibited only mild hypointensities likely associated with hemorrhage on T2WI and T2*WI.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3585319&req=5

pone-0056125-g007: A demonstration in vivo cellular MRI using the simple labeling method.A rat with glioma was transplanted with SPION-labeled NPCs into the lateral ventricle, and T2WIs were used to track the migration and distribution of the transplanted NPCs. (A) T2WI acquired at baseline, on day 1, 7, and 14 after transplantation. Hypointensities were readily visible one day later, and persisted throughout the tracking duration. (B) Signals at the tumor or injection site in relation to a control area (the cortex) were plotted against time. (C) PB staining confirmed the relocation of SPION-labeled cells following transplantation. Histology was performed on brain tissues obtained on day 14 after transplantation. The white arrow indicates the location of the tumor whereas the red arrows indicate the hypointensity arising from the labeled NPCs. For tumors without injected NPCs, the signal patterns were different. Untreated ENU tumors exhibited only mild hypointensities likely associated with hemorrhage on T2WI and T2*WI.
Mentions: The T2*WI and T2WI at baseline and at 1, 7, and 14 days after transplantation of SPION-labeled NPCs into the lateral ventricle of the rat with glioma are shown in Fig. 7A. The tumor was identified as a hyperintensity at baseline. One day following transplantation, the signal at the tumor site became hypointense and persisted throughout the tracking duration. Quantification of the signal at the tumor or injection site over time is shown in Fig. 7B. The tumor exhibited increasing hypointensity and the injection site remained the same hypointense. PB staining performed on the brain tissues obtained 14 days after transplantation indicated that iron was present in the tumor site, which should be responsible for the signal changes on T2*WI and T2WI (Fig. 7C). For tumors without injected NPCs, the signal patterns were different. Untreated ENU tumors exhibited only mild hypointensities associated with hemorrhage on T2WI and T2*WI (data not shown).

Bottom Line: However, compelling evidence also shows that simple SPION incubation is not invariably ineffective.The labeling efficiency can be improved by prolonged incubation and elevated iron doses.The results showed that, following 48 hours of incubation at 75 µg/ml, nearly all NPCs exhibited visible SPION intake.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biomedical Sciences, Academic Sinica, Taipei, Taiwan.

ABSTRACT
Cellular magnetic resonance imaging (MRI) has been well-established for tracking neural progenitor cells (NPC). Superparamagnetic iron oxide nanoparticles (SPIONs) approved for clinical application are the most common agents used for labeling. Conventionally, transfection agents (TAs) were added with SPIONs to facilitate cell labeling because SPIONs in the native unmodified form were deemed inefficient for intracellular labeling. However, compelling evidence also shows that simple SPION incubation is not invariably ineffective. The labeling efficiency can be improved by prolonged incubation and elevated iron doses. The goal of the present study was to establish simple SPION incubation as an efficient intracellular labeling method. To this end, NPCs derived from the neonatal subventricular zone were incubated with SPIONs (Feridex®) and then evaluated in vitro with regard to the labeling efficiency and biological functions. The results showed that, following 48 hours of incubation at 75 µg/ml, nearly all NPCs exhibited visible SPION intake. Evidence from light microscopy, electron microscopy, chemical analysis, and magnetic resonance imaging confirmed the effectiveness of the labeling. Additionally, biological assays showed that the labeled NPCs exhibited unaffected viability, oxidative stress, apoptosis and differentiation. In the demonstrated in vivo cellular MRI experiment, the hypointensities representing the SPION labeled NPCs remained observable throughout the entire tracking period. The findings indicate that simple SPION incubation without the addition of TAs is an efficient intracellular magnetic labeling method. This simple approach may be considered as an alternative approach to the mainstream labeling method that involves the use of TAs.

Show MeSH
Related in: MedlinePlus