Region-specific differentiation of the hippocampal stem cell line HiB5 upon implantation into the developing mammalian brain

Evaluation of Tet-on system to avoid transgene down-regulation in ex vivo gene transfer to the CNS

“…Previous in vivo studies have shown that the immortalized HiB5 cell line survive grafting very well, remain stable for at least 6 months and become structurally integrated within an area surrounding the implantation site. [13][14][15] The in vivo expression stability of UbC and CMV constitutive promoters was compared by transplanting HiB5 cells stably expressing EGFP from the UbC promoter or the CMV promoter to the striatum. Expression of EGFP from both promoters was significantly down-regulated over 21 days ( Figure 2) and no difference in expression stability was seen between cells expressing EGFP under the CMV or UbC promoters, respectively (data from CMV-EGFP not shown).…”

“…13 Cells were expanded at the permissive temperature for the immortalizing oncogene (+33°C, tsA58/U19 mutant allele of the SV40 large T-antigen) in Dulbecco's modified Eagle's medium with Glutamax (Gibco, Life Technologies, Denmark) supplemented with 10% heatinactivated fetal bovine serum and 5 g/ml gentamycin. Plasmid transfections in HiB5 cells or CHO cells were done using the Lipo Plus transfection system in accordance with the manufacturer's (Life Technologies) recommendations.…”

Gene transfer to the central nervous system by transplantation of cerebral endothelial cells

“…The transplantation of immortalized neural Serotec, Oxford, UK), Glut-1 (Biogenesis, Bournemouth, UK), rat laminin (Sigma), GFAP (Dako, Glostrup, progenitor cells in the brain has also been studied recently. [2][3][4] Those articles described the absence of Denmark). The specificity of peroxidase and fluorescent immunolabelling was confirmed by the absence of staintumorigenicity of such cell lines and their integration and differentiation capacity in vivo.…”

“…17,18 When functional integration of neural progenitor cell lines into co-implanted in rat brains with different types of glioma host CNS. [2][3][4] These data, together with the observation cells, ␤-galactosidase expressing RBE4 cells (RBEZ cells) that vascular endothelial cells were efficiently used as were shown to survive for up to 5 weeks and to be gene carriers in peripheral organs, [5][6][7][8][9][10][11][12] prompted us to frequently associated with tumor-infiltrating vascular evaluate the brain endothelial cell as an alternative route profiles. 20 for gene transfer to the CNS.…”

Truncated N-terminal mutants of SV40 large T antigen as minimal immortalizing agents for CNS cells

“…Cells immortalized with LT, either wild type or temperature sensitive variants, typically are not tumorigenic after transplantation into the brain (Giordano et al, 1996;Isono et al, 1992;Okoye et al, 1994Okoye et al, ,1995Renfranz et al, 1991), but are often phenotypically immature and genetically abnormal (Cepko, 1992); reviewed in Bryan and Reddel (1994).…”