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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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Multiple differentiation of NSCs following AAV transduction (A–D) NSCs differentiated into various cell types with various morphological appearances following 7 days in differentiation medium. Immunocytochemistry further confirmed that these cells were: GFAP-positive astrocytes (E) and β-tubullin III-positive neurons (F). E,F Scal bar, 100 um
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f3-medscimonit-17-11-br305: Multiple differentiation of NSCs following AAV transduction (A–D) NSCs differentiated into various cell types with various morphological appearances following 7 days in differentiation medium. Immunocytochemistry further confirmed that these cells were: GFAP-positive astrocytes (E) and β-tubullin III-positive neurons (F). E,F Scal bar, 100 um

Mentions: The ability of NSCs to form neurospheres and undergo differentiation was determined following AAV transduction. In both the AAV-NT-3- and AAV-GFP-transduced groups, abundant spherical cell clusters emerged after 5–7 d in growth medium. After seeding in differentiation medium, cells in both groups differentiated into various cell types as defined by different morphological appearance (Figure 3A–D). Immunocytochemistry further confirmed that the differentiated cells were GFAP-positive astrocytes and β-tubulin III-positive neurons (Figure 3E,F).


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)

Multiple differentiation of NSCs following AAV transduction (A–D) NSCs differentiated into various cell types with various morphological appearances following 7 days in differentiation medium. Immunocytochemistry further confirmed that these cells were: GFAP-positive astrocytes (E) and β-tubullin III-positive neurons (F). E,F Scal bar, 100 um
© Copyright Policy
Related In: Results  -  Collection

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

f3-medscimonit-17-11-br305: Multiple differentiation of NSCs following AAV transduction (A–D) NSCs differentiated into various cell types with various morphological appearances following 7 days in differentiation medium. Immunocytochemistry further confirmed that these cells were: GFAP-positive astrocytes (E) and β-tubullin III-positive neurons (F). E,F Scal bar, 100 um
Mentions: The ability of NSCs to form neurospheres and undergo differentiation was determined following AAV transduction. In both the AAV-NT-3- and AAV-GFP-transduced groups, abundant spherical cell clusters emerged after 5–7 d in growth medium. After seeding in differentiation medium, cells in both groups differentiated into various cell types as defined by different morphological appearance (Figure 3A–D). Immunocytochemistry further confirmed that the differentiated cells were GFAP-positive astrocytes and β-tubulin III-positive neurons (Figure 3E,F).

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