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Human Gene Therapy
Characterization of 911: A New Helper Cell Line for the Titration and Propagation of Early Region 1-Deleted Adenoviral Vectors
To cite this article:
Frits J. Fallaux, Onno Kranenburg, Steve J. Cramer, Ada Houweling, Hans van Ormondt, Rob C. Hoeben, Alex J. van der Eb.
Human Gene Therapy.
January 1996,
7(2): 215-222.
doi:10.1089/hum.1996.7.2-215.
Address reprint requests to: Prof. Alex J. van der Eb, Department of Medical Biochemistry, Sylvius Laboratories, PO Box 9503, 2300 RA Leiden, The Netherlands 03 03 1995 Received for publication March 3, 1995. 21 09 1995 Accepted after revision September 21, 1995. ABSTRACT Currently, the preferred host for the production of early region-1 (E1)-deleted recombinant adenoviruses (rAdV) is cell line 293, which was generated by transformation of human embryonic kidney cells by sheared adenovirus 5 (Ad5) DNA. To develop alternative hosts for the production of rAdV, we generated adenovirus-transformed human cell lines by transformation of human embryonic retinoblasts (HER) with a plasmid containing base pairs 79–5789 of the Ad5 genome. One of the established HER cell lines, which we called 911, exhibited favorable growth characteristics and was chosen for further study. This cell line is demonstrated to have several characteristics in common with the well-known 293 cell line: The 911 cell line is highly transfectable, and exhibits similar frequencies of homologous recombination. However, it has additional characteristics that make it a useful alternative for 293. The 911 cells perform particularly well in plaque assays. Upon infection with E1-deleted adenoviruses, plaques become apparent in monolayers of 911 cells already after 3–4 days versus 4–10 days in monolayers of 293 cells, thereby reducing the time required for quantitative plaque assays. Furthermore, yields of E1-deleted adenovirus vectors up to three times as high as those achieved with 293 cells can be obtained with 911 cells. Finally, the Ad5-DNA content of the 911 cell line is completely known. These features make the 911 cell line a useful alternative for the construction, propagation, and titration of E1-deleted recombinant adenoviruses. Overview summary The recombinant adenoviruses used today as gene transfer vectors for gene therapy purposes have a deletion in the E1 region of the viral genome. The defective gene products are provided in helper cells, thus allowing the defective viruses to be propagated. At the moment, there is only one widely available E1-expressing helper cell line, 293. This study describes the characterization of a new E1-complementing cell line 911 established by transformation of human embryonic retinoblasts with Ad5-E1. The authors demonstrate that 911 cells perform especially well in plaque assays and in small-scale production of adenovirus vectors.  This paper was cited by:E. coli NfsA: an alternative nitroreductase for prodrug activation gene therapy in combination with CB1954 S O Vass, D Jarrom, W R Wilson, E I Hyde, P F Searle British Journal of Cancer. Jul 2009, Vol. 100, No. 12: 1903-1911 CrossRef Comparison of Viral and Nonviral Vectors for Gene Transfer to Human Endothelial Progenitor Cells Brian Kealy, Aaron Liew, Jill M. McMahon, Thomas Ritter, Aideen O'Doherty, Melissa Hoare, Udo Greiser, Erin E. Vaughan, Martin Maenz, Ciara O'Shea, Frank Barry, Timothy O'Brien Tissue Engineering Part C: Methods. Jun 2009, Vol. 15, No. 2: 223-231 Abstract | Full Text PDF or HTML | Reprints & PermissionsA strategy for genetic modification of the spike-encoding segment of human reovirus T3D for reovirus targeting D J M van den Wollenberg, S K van den Hengel, I J C Dautzenberg, S J Cramer, O Kranenburg, R C Hoeben Gene Therapy. Jan 2009, Vol. 15, No. 24: 1567-1578 CrossRef Gene Transfer May Be Preventive But Not Curative for a Lysosomal Transport Disorder Claire Hippert, Grégor Dubois, Carole Morin, Olivier Disson, Sandy Ibanes, Chantal Jacquet, Reto Schwendener, Corinne Antignac, Eric J Kremer, Vasiliki Kalatzis Molecular Therapy. Sep 2008, Vol. 16, No. 8: 1372-1381 CrossRef CD40ligand‐expressing dendritic cells induce regression of hepatocellular carcinoma by activating innate and acquired immunity in vivo Maria A. Gonzalez‐Carmona, Veronika Lukacs‐Kornek, Anne Timmerman, Sara Shabani, Miroslaw Kornek, Annabelle Vogt, Yildiz Yildiz, Elisabeth Sievers, Ingo G.H. Schmidt‐Wolf, Wolfgang H. Caselmann, Tilman Sauerbruch, Volker Schmitz Hepatology. Aug 2008, Vol. 48, No. 1: 157-168 CrossRef Adenovirus targeting to HLA-A1/MAGE-A1-positive tumor cells by fusing a single-chain T-cell receptor with minor capsid protein IX J de Vrij, T G Uil, S K van den Hengel, S J Cramer, D Koppers-Lalic, M C Verweij, E J H J Wiertz, J Vellinga, R A Willemsen, R C Hoeben Gene Therapy. Aug 2008, Vol. 15, No. 13: 978-989 CrossRef Serum-free transient protein production system based on adenoviral vector and PER.C6 technology: High yield and preserved bioactivity M.J.E. Havenga, L. Holterman, I. Melis, S. Smits, J. Kaspers, E. Heemskerk, R. van der Vlugt, M. Koldijk, G.J. Schouten, G. Hateboer, K. Brouwer, R. Vogels, J. Goudsmit Biotechnology and Bioengineering. Jul 2008, Vol. 100, No. 2: 273-283 CrossRef Cytokine-induced β-cell apoptosis is NO-dependent, mitochondria-mediated and inhibited by BCL-X
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