title: Retroviral transfection of the lacZ gene from Liz-9 packaging cells to glioma spheroids.
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 beta-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.
authors.affiliation: Department of Neurosurgery, Haukeland Hospital, Bergen, Norway.
authors.forename: P E
authors.initials: PE
authors.lastname: Oyvind
authors.forename: T
authors.initials: T
authors.lastname: Visted
authors.forename: F
authors.initials: F
authors.lastname: Thorsen
authors.forename: R
authors.initials: R
authors.lastname: Bjerkvig
authors.forename: M
authors.initials: M
authors.lastname: Lund-Johansen
mesh_terms: D001932:Brain Neoplasms / Q000628:therapy*; D002460:Cell Line; D015316:Genetic Therapy; D005822:Genetic Vectors; D005910:Glioma / Q000628:therapy*; D006801:Humans; D007763:Lac Operon; D012190:Retroviridae / Q000235:genetics*; D018874:Spheroids, Cellular; D014162:Transfection; D014407:Tumor Cells, Cultured