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Neurotrophin-3 gene transduction of mouse neural stem cells promotes proliferation and neuronal differentiation in organotypic hippocampal slice cultures.

Lu HX, Hao ZM, Jiao Q, Xie WL, Zhang JF, Lu YF, Cai M, Wang YY, Yang ZQ, Parker T, Liu Y - Med. Sci. Monit. (2011)

Bottom Line: The effect of NT-3 over-expression on cell proliferation and differentiation in NSCs was observed by immunohistochemistry, cell culture and organotypic hippocampal slice cultures.<br /> The characteristics of self-renewal and multiple differentiation of NSCs were well-preserved.Furthermore, cells in the NSC-NT-3 group survived in a significantly higher percentage and undertook neuronal differentiation preferably in organotypic hippocampal slice cultures.Our results suggest that the transduction of NT-3 into NSCs could effectively promote NSCs survival, proliferation, and neuronal differentiation in vitro without change of the stemness of NSCs.

View Article: PubMed Central - PubMed

Affiliation: Institute of Neurobiology, Environment and Genes Related to Diseases Key Laboratory of Education Ministry, Xi'an Jiaotong University College of Medicine, Xi'an, PR China.

ABSTRACT

Background: The transplantation of neural stem cells (NSCs) has been accepted as a promising therapeutic strategy for central nervous system disorders. However, the beneficial effect of NSC transplantation upon functional recovery is limited due to the unfavorable microenvironment (niche) at the site of trauma or degenerative disease in the brain. Combination of transplantation of NSCs with neurotrophins may overcome the hurdles of impaired cell survival and neuronal differentiation.

Material/methods: In the current study, the neurotrophin-3 (NT-3) gene was transduced into cultured mouse embryonic cortical NSCs via an AAV vector (NSC-NT-3). The effect of NT-3 over-expression on cell proliferation and differentiation in NSCs was observed by immunohistochemistry, cell culture and organotypic hippocampal slice cultures.

Results: The characteristics of self-renewal and multiple differentiation of NSCs were well-preserved. Cells in the NSC-NT-3 group proliferated faster and differentiated into more β-tubulin III-positive neurons compared to the control group in vitro. Furthermore, cells in the NSC-NT-3 group survived in a significantly higher percentage and undertook neuronal differentiation preferably in organotypic hippocampal slice cultures.

Conclusions: Our results suggest that the transduction of NT-3 into NSCs could effectively promote NSCs survival, proliferation, and neuronal differentiation in vitro without change of the stemness of NSCs. This work also offers evidence to better understand the safety and efficiency of combined treatment with NT-3 and NSCs for the central nervous system disorders.

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Related in: MedlinePlus

The survival and differentiation of NSCs-NT-3 in orgnaotypic hippocampal slice cultures Hippocampal slice cultures remained healthy after addition of CM-DiI labeled neurospheres (A). CM-DiI labeled neurospheres (insert in B) appeared to all over the slices and more NSCs-NT-3 survived after transplanted into organotypic hippocampal slices than controls (B). Some of them differentiated into β-tubulin positive neurons (C) and GFAP positive astrocytes (D). A Scal bar, 100 um; B Scal bar, 50 um; C and D Scal bar, 20 um.
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f5-medscimonit-17-11-br305: The survival and differentiation of NSCs-NT-3 in orgnaotypic hippocampal slice cultures Hippocampal slice cultures remained healthy after addition of CM-DiI labeled neurospheres (A). CM-DiI labeled neurospheres (insert in B) appeared to all over the slices and more NSCs-NT-3 survived after transplanted into organotypic hippocampal slices than controls (B). Some of them differentiated into β-tubulin positive neurons (C) and GFAP positive astrocytes (D). A Scal bar, 100 um; B Scal bar, 50 um; C and D Scal bar, 20 um.

Mentions: All slices remained healthy after addition of CM-DiI labeled neurospheres (Figure 5A). CM-DiI labeled NSCs appeared throughout the slices. It was noticed that more NSCs-NT-3 survived after being transplanted into organotypic hippocampal slices than controls (Figure 5B). After 4 days of co-culture, abundant GFAP-positive glia and β-tubulin III positive neurons were observed throughout the slices. Some of them were labeled with both CM-DiI and β-tubulin or CM-DiI and GFAPs (Figure 5C, D), indicating the differentiation of transplanted NSCs in slice cultures. The proportion of neuron and glia differentiation was similar to that observed in vitro.


Neurotrophin-3 gene transduction of mouse neural stem cells promotes proliferation and neuronal differentiation in organotypic hippocampal slice cultures.

Lu HX, Hao ZM, Jiao Q, Xie WL, Zhang JF, Lu YF, Cai M, Wang YY, Yang ZQ, Parker T, Liu Y - Med. Sci. Monit. (2011)

The survival and differentiation of NSCs-NT-3 in orgnaotypic hippocampal slice cultures Hippocampal slice cultures remained healthy after addition of CM-DiI labeled neurospheres (A). CM-DiI labeled neurospheres (insert in B) appeared to all over the slices and more NSCs-NT-3 survived after transplanted into organotypic hippocampal slices than controls (B). Some of them differentiated into β-tubulin positive neurons (C) and GFAP positive astrocytes (D). A Scal bar, 100 um; B Scal bar, 50 um; C and D Scal bar, 20 um.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3539508&req=5

f5-medscimonit-17-11-br305: The survival and differentiation of NSCs-NT-3 in orgnaotypic hippocampal slice cultures Hippocampal slice cultures remained healthy after addition of CM-DiI labeled neurospheres (A). CM-DiI labeled neurospheres (insert in B) appeared to all over the slices and more NSCs-NT-3 survived after transplanted into organotypic hippocampal slices than controls (B). Some of them differentiated into β-tubulin positive neurons (C) and GFAP positive astrocytes (D). A Scal bar, 100 um; B Scal bar, 50 um; C and D Scal bar, 20 um.
Mentions: All slices remained healthy after addition of CM-DiI labeled neurospheres (Figure 5A). CM-DiI labeled NSCs appeared throughout the slices. It was noticed that more NSCs-NT-3 survived after being transplanted into organotypic hippocampal slices than controls (Figure 5B). After 4 days of co-culture, abundant GFAP-positive glia and β-tubulin III positive neurons were observed throughout the slices. Some of them were labeled with both CM-DiI and β-tubulin or CM-DiI and GFAPs (Figure 5C, D), indicating the differentiation of transplanted NSCs in slice cultures. The proportion of neuron and glia differentiation was similar to that observed in vitro.

Bottom Line: The effect of NT-3 over-expression on cell proliferation and differentiation in NSCs was observed by immunohistochemistry, cell culture and organotypic hippocampal slice cultures.<br /> The characteristics of self-renewal and multiple differentiation of NSCs were well-preserved.Furthermore, cells in the NSC-NT-3 group survived in a significantly higher percentage and undertook neuronal differentiation preferably in organotypic hippocampal slice cultures.Our results suggest that the transduction of NT-3 into NSCs could effectively promote NSCs survival, proliferation, and neuronal differentiation in vitro without change of the stemness of NSCs.

View Article: PubMed Central - PubMed

Affiliation: Institute of Neurobiology, Environment and Genes Related to Diseases Key Laboratory of Education Ministry, Xi'an Jiaotong University College of Medicine, Xi'an, PR China.

ABSTRACT

Background: The transplantation of neural stem cells (NSCs) has been accepted as a promising therapeutic strategy for central nervous system disorders. However, the beneficial effect of NSC transplantation upon functional recovery is limited due to the unfavorable microenvironment (niche) at the site of trauma or degenerative disease in the brain. Combination of transplantation of NSCs with neurotrophins may overcome the hurdles of impaired cell survival and neuronal differentiation.

Material/methods: In the current study, the neurotrophin-3 (NT-3) gene was transduced into cultured mouse embryonic cortical NSCs via an AAV vector (NSC-NT-3). The effect of NT-3 over-expression on cell proliferation and differentiation in NSCs was observed by immunohistochemistry, cell culture and organotypic hippocampal slice cultures.

Results: The characteristics of self-renewal and multiple differentiation of NSCs were well-preserved. Cells in the NSC-NT-3 group proliferated faster and differentiated into more β-tubulin III-positive neurons compared to the control group in vitro. Furthermore, cells in the NSC-NT-3 group survived in a significantly higher percentage and undertook neuronal differentiation preferably in organotypic hippocampal slice cultures.

Conclusions: Our results suggest that the transduction of NT-3 into NSCs could effectively promote NSCs survival, proliferation, and neuronal differentiation in vitro without change of the stemness of NSCs. This work also offers evidence to better understand the safety and efficiency of combined treatment with NT-3 and NSCs for the central nervous system disorders.

Show MeSH
Related in: MedlinePlus