These authors contributed equally to this publication.
A retroviral packaging cell line for pseudotype vectors based on glioma-infiltrating progenitor cells
Version of Record online: 1 MAY 2007
Copyright © 2007 John Wiley & Sons, Ltd.
The Journal of Gene Medicine
Volume 9, Issue 5, pages 335–344, May 2007
How to Cite
Fischer, Y. H., Miletic, H., Giroglou, T., Litwak, S., Stenzel, W., Neumann, H. and von Laer, D. (2007), A retroviral packaging cell line for pseudotype vectors based on glioma-infiltrating progenitor cells. J. Gene Med., 9: 335–344. doi: 10.1002/jgm.1032
- Issue online: 1 MAY 2007
- Version of Record online: 1 MAY 2007
- Manuscript Accepted: 19 FEB 2007
- Manuscript Revised: 5 FEB 2007
- Manuscript Received: 6 OCT 2006
- Hertie-Foundation and Walter-und-Ilse-Rose-Foundation
- Köln Fortune Program. Grant Number: 108/2003
Early clinical trials for gene therapy of human gliomas with retroviral packaging cells (PC) have been hampered by low transduction efficacy and lack of dissemination of PC within the tumor. In the current approach, these issues have been addressed by creating a stable packaging cell line for retroviral vectors pseudotyped with glycoproteins of lymphocytic choriomeningitis virus (LCMV) based on tumor-infiltrating progenitor cells.
Tumor-infiltrating progenitor cells, which had been isolated from adult rat bone marrow (BM-TIC), were modified to stably express Gag-Pol proteins of moloney murine leukemia virus (Mo-MLV) and glycoproteins of LCMV. Packaging of a retroviral vector was measured by titration experiments on human fibroblast cells as well as on mouse and human glioma cell lines. Additionally, gene transfer was tested in a rat glioma model in vivo.
The BM-TIC-derived packaging cell line (BM-TIPC) produced retroviral vectors with titers between 2–8 × 103 transducing units (TU)/ml. Extended culturing of BM-TIPC over several weeks and freezing/thawing of cells did not affect vector titers. No replication-competent retrovirus was released from BM-TIPC. In a rat glioma model, BM-TIPC infiltrated the tumors extensively and with high specificity. Moreover, BM-TIPC mediated transduction of glioma cells in vivo.
This proof-of-principle study shows that primary adult progenitor cells with tumor-infiltrating capacity can be genetically modified to stably produce retroviral LCMV pseudotype vectors. These BM-TIPC may be a useful tool to enhance specificity and efficacy of gene transfer to gliomas in patients. Copyright © 2007 John Wiley & Sons, Ltd.