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Effective adenovirus-mediated gene transfer into neural stem cells derived from human embryonic stem cells

Journal Article


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 (∼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.

Authors


  •   Bertram, Cornelia (external author)
  •   Hawes, Susan M. (external author)
  •   Egli, Simone (external author)
  •   Peh, Gary (external author)
  •   Dottori, Mirella
  •   Kees, Ursula (external author)
  •   Dallas, Peter (external author)

Publication Date


  • 2010

Citation


  • Bertram, C. M., Hawes, S. M., Egli, S., Peh, G. S. L., Dottori, M., Kees, U. R. & Dallas, P. B. (2010). Effective adenovirus-mediated gene transfer into neural stem cells derived from human embryonic stem cells. Stem Cells and Development, 19 (4), 569-577.

Scopus Eid


  • 2-s2.0-77950618467

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/1177

Has Global Citation Frequency


Number Of Pages


  • 8

Start Page


  • 569

End Page


  • 577

Volume


  • 19

Issue


  • 4

Place Of Publication


  • United States

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 (∼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.

Authors


  •   Bertram, Cornelia (external author)
  •   Hawes, Susan M. (external author)
  •   Egli, Simone (external author)
  •   Peh, Gary (external author)
  •   Dottori, Mirella
  •   Kees, Ursula (external author)
  •   Dallas, Peter (external author)

Publication Date


  • 2010

Citation


  • Bertram, C. M., Hawes, S. M., Egli, S., Peh, G. S. L., Dottori, M., Kees, U. R. & Dallas, P. B. (2010). Effective adenovirus-mediated gene transfer into neural stem cells derived from human embryonic stem cells. Stem Cells and Development, 19 (4), 569-577.

Scopus Eid


  • 2-s2.0-77950618467

Ro Metadata Url


  • http://ro.uow.edu.au/ihmri/1177

Has Global Citation Frequency


Number Of Pages


  • 8

Start Page


  • 569

End Page


  • 577

Volume


  • 19

Issue


  • 4

Place Of Publication


  • United States