Effective adenovirus-mediated gene transfer into neural stem cells derived from human embryonic stem cells.
Stem Cells Dev
; 19(4): 569-78, 2010 Apr.
Article
em En
| MEDLINE
| ID: mdl-19594361
ABSTRACT
Human embryonic stem cell-derived neural stem cells (hESC-NSCs) are an attractive cell type for studying aspects of brain development and pathology. To develop the full potential of this model system, it is important to establish a reliable methodology for the manipulation of gene expression in hNSCs. To address this issue, we used an adenoviral vector with a CMV promoter-driven green fluorescent protein (GFP) reporter gene (Ad5-GFP). We optimized conditions for Ad5-GFP infection and assessed the efficiency of infection of whole and dissociated embryonic stem cell (ESC)-derived neurospheres as well as the effect of adenoviral vectors on cell surface marker expression, proliferation, and differentiation potential. Our results demonstrate that most neurosphere cells ( approximately 70%) express the coxsackie and adenovirus receptor and can be infected with Ad5. More specifically, the CD133+ hESC-NSC population could be infected more efficiently than the CD133 population and both populations expressed GFP at high levels. At low multiplicity of infection (MOI < 25), the virus had no significant effect on stem cell marker expression (CD133 and Nestin), cell survival, cell proliferation rate, or differentiation potential. This model system provides a practical new approach to study human NSC function in the context of neurodegenerative and neoplastic disorders.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Transdução Genética
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Adenoviridae
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Técnicas de Transferência de Genes
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Células-Tronco Embrionárias
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Vetores Genéticos
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Neurônios
Limite:
Humans
Idioma:
En
Ano de publicação:
2010
Tipo de documento:
Article