Retroviral transfection of the lacZ gene from Liz-9 packaging cells to glioma spheroids

Per Enger Øyvind, Therese Visted*, Frits Thorsen, Rolf Bjerkvig, Morten Lund-Johansen

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

7 Citations (Scopus)

Abstract

Despite the development of numerous vectors for gene transfection to gliomas, patient survival length remains unaffected in clinical trials. For glioma gene therapy to be successful, the extent of gene transfer to the solid tumor tissue has to be high. In the present work we review some of the vector types and strategies so far utilized in experimental and clinical glioma gene therapy. Since gene transfer efficacy into solid glioma tissue is unknown for many vectors, we studied the gene transfer efficacy into multicellular spheroids derived from a human glioma cell line GaMg as well as into spheroids derived from human glioma biopsies (glioblastoma multiforme, GBM). A replication deficient retroviral vector from the Liz 9 packaging cell line was used for transfer of the bacterial β-galactosidase lacZ gene into the target tissue. Gene transfer was obtained by adding medium containing virus from the producer cells to the target tissue. The experiments were also conducted with EGF (epidermal growth factor) added to the medium. The data show that the transfection rate ranged from 0-4.5% where the transfection efficacy was higher in spheroids after the addition of EGF. Most of the transfected cells were found at the surface, but transfected cells could also be observed in the center of the spheroids. We conclude that using this vector system, the transfection efficacy was low, even if the number of replicating cells was increased by adding EGF. The findings are consistent, and may partly explain, the lack of effect using this vector system during in vivo studies. Copyright (C) 1999 ISDN.

Original languageEnglish
Pages (from-to)665-672
Number of pages8
JournalInternational Journal of Developmental Neuroscience
Volume17
Issue number5-6
DOIs
Publication statusPublished - Aug 1999
Externally publishedYes

Keywords

  • GaMg
  • Gene therapy
  • Glioblastoma multiforme
  • Multicellular spheroids
  • Retroviral transfection

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