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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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SPION-labeled NPCs formed neurospheres.(A) The neural progenitor identify confirmed by nestin immunostaining. (B) Nuclear staining by DAPI. (C) Merged nestin and DAPI staining. The majority of the cells within the sphere were nestin positive. (D) SPION labeling was examined by PB staining with nuclear fast red counterstain. (E) An enlarged view of dissociated SPION labeled NPCs stained with PB.
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pone-0056125-g002: SPION-labeled NPCs formed neurospheres.(A) The neural progenitor identify confirmed by nestin immunostaining. (B) Nuclear staining by DAPI. (C) Merged nestin and DAPI staining. The majority of the cells within the sphere were nestin positive. (D) SPION labeling was examined by PB staining with nuclear fast red counterstain. (E) An enlarged view of dissociated SPION labeled NPCs stained with PB.

Mentions: The SPION labeled NPCs formed neurospheres. The neurospheres formed by labeled NPCs coincided with those by unlabeled NPCs in the size (∼150 uM), shape (spherical), and time (∼7 days). Nestin immunostaining (Fig. 2A), DAPI (Fig. 2B), and the merged image (Fig. 2C) indicate the neural progenitor identity of the cells. The neurospheres were stained positive for PB (Fig. 2D). An enlarged view of the dissociated SPION labeled NPCs is shown in Fig. 2E.


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)

SPION-labeled NPCs formed neurospheres.(A) The neural progenitor identify confirmed by nestin immunostaining. (B) Nuclear staining by DAPI. (C) Merged nestin and DAPI staining. The majority of the cells within the sphere were nestin positive. (D) SPION labeling was examined by PB staining with nuclear fast red counterstain. (E) An enlarged view of dissociated SPION labeled NPCs stained with PB.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585319&req=5

pone-0056125-g002: SPION-labeled NPCs formed neurospheres.(A) The neural progenitor identify confirmed by nestin immunostaining. (B) Nuclear staining by DAPI. (C) Merged nestin and DAPI staining. The majority of the cells within the sphere were nestin positive. (D) SPION labeling was examined by PB staining with nuclear fast red counterstain. (E) An enlarged view of dissociated SPION labeled NPCs stained with PB.
Mentions: The SPION labeled NPCs formed neurospheres. The neurospheres formed by labeled NPCs coincided with those by unlabeled NPCs in the size (∼150 uM), shape (spherical), and time (∼7 days). Nestin immunostaining (Fig. 2A), DAPI (Fig. 2B), and the merged image (Fig. 2C) indicate the neural progenitor identity of the cells. The neurospheres were stained positive for PB (Fig. 2D). An enlarged view of the dissociated SPION labeled NPCs is shown in Fig. 2E.

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