Clin Exp Otorhinolaryngol.  2010 Jun;3(2):84-90. 10.3342/ceo.2010.3.2.84.

Transplantation of Neural Stem Cells in Anosmic Mice

• Lee CH
• Jeon SW
• Seo BS
• Mo JH
• Jeon EH
• Choi AR
• Kim JW

• KMID: 1460594
• DOI: http://doi.org/10.3342/ceo.2010.3.2.84

Abstract

OBJECTIVES
Treating olfactory dysfunction is a challenge for physicians. One of the therapeutic options could be transplantation of stem cells. In this study, neural stem cells were transplanted into anosmic mice.
METHODS
Neural stem cells were generated from the olfactory bulb of green fluorescent protein (GFP)-transgenic C57BL6 mice. Anosmia were induced by injection of intraperitoneal 3-methylindole. The neural stem cells were transplanted transnasally on the next day. The olfactory function was evaluated by a food-finding test once a week. The olfactory neuroepithelium was harvested for histologic examination and protein analysis at 4 weeks.
RESULTS
Twenty-five percent (6/24) of the control mice that were not transplanted with neural stem cells survived at 4 weeks while 67% (8/12) of the transplanted mice survived (P=0.029). The food finding test showed that the transplanted mice resumed finding food at 3 weeks while the control mice resumed finding food at 4 weeks. GFP-positive cells were observed in the olfactory neuroepithelium of the transplanted mice. Western blotting revealed that the olfactory marker protein expression was significantly lower in the control mice than that in the transplanted mice.
CONCLUSION
This study demonstrated that improvement of mouse survival was achieved and recovery of olfactory function was promoted by transnasal transplantation of neural stem cells in the anosmic mouse model. These results indicate that stem cells might be one of the future modalities for treating olfactory impairment.

Keyword

Neural stem cell; Anosmia; Transplantation; Smell

MeSH Terms

Animals
Blotting, Western
Mice
Neural Stem Cells
Olfaction Disorders
Olfactory Bulb
Olfactory Marker Protein
Skatole
Smell
Stem Cells
Transplants
Olfactory Marker Protein
Skatole

Figure

• 

  Fig. 1 Neurospheres from the olfactory bulb of the green fluorescence protein-transgenic C57BL6 mice (A). The neural stem cells have formed neurospheres floating in the culture media. Immunoreactivity to a marker for neural stem cells (B). The neurosphere-forming cells were immunoreactive to nestin (red), which is a neural stem cell marker.

• 

  Fig. 2 The survival analysis. Six out of twenty-four control mice that were not transplanted with neural stem cells (NSC) survived at 4 weeks after injection of 3-methylindole, while 8 out of the 12 transplanted mice survived at 4 weeks (P=0.029).

• 

  Fig. 3 The olfactory function. The olfactory function was evaluated by the time spent to find a piece of cheese buried beneath the wood shavings. Five out of six transplanted mice (right) with neural stem cells (NSC) could find the food at 3 weeks while two out of six control mice could find the food at 3 weeks (left). At 3 weeks, there was no significant difference in the time spent to find the cheese between the control and transplanted mice (P=0.13). On the other hand, at 4 weeks, the cheese was found more quickly by the transplanted mice than by the control mice (P=0.04). NSC (-) denotes without transplantation of NSC while NSC (+) with transplantation.

• 

  Fig. 4 Neural stem cells in the whole-mount olfactory neuroepithelium. The whole tissue mounting showed several conglomerates of green fluorescent neural stem cells that had been transplanted and they resided in the olfactory neuroepithlium.

• 

  Fig. 5 Immunohistochemical staining for olfactory marker protein (OMP) and green fluorescent protein (GFP). The staining was performed on the consecutive sections of the olfactory turbinates. OMP-positive cells were observed (A) in the olfactory neuroepithelium from the healthy wild-type mice that were not treated with 3-mehtylindole and that had normal olfactory function, while there were no GFP-positive cells (B). There were no OMP- (C) or GFP-positive cells (D) in the olfactory neuroepithelium from the anosmic mice that were not transplanted with neural stem cells. In the transplanted mice, the cells were only positive for OMP (E) but not for GFP (F) in some olfactory neuroepithelial regions, whereas cells that were positive for both OMP (G) and GFP (H) were observed in some regions.

• 

  Fig. 6 Western immunoblotting of the olfactory neuroepithelium. Olfactory marker protein (OMP) was more highly expressed in the transplanted mice than that in the control mice (A). Densitometric analysis showed a significant difference of the OMP expression between the control and transplanted mice (B). neural stem cells (NSC) (-) denotes without transplantation of neural stem cells while NSC (+) with transplantation.

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