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By Ke Xu

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Optimized conditions for the production of recombinant amphotropic retroviral vector preparations. Gene Ther 4(2):172-6. Kotani H, Newton PB, 3rd, Zhang S, Chiang YL, Otto E, Weaver L, Blaese RM, Anderson WF, McGarrity GJ. 1994. Improved methods of retroviral vector transduction and production for gene therapy. Hum Gene Ther 5(1):19-28. Kotsopoulou E, Kim VN, Kingsman AJ, Kingsman SM, Mitrophanous KA. 2000. A Revindependent human immunodeficiency virus type 1 (HIV-1)-based vector that exploits a codon-optimized HIV-1 gag-pol gene.

Gene Ther 15(18):1280-6. Lewis P, Hensel M, Emerman M. 1992. Human immunodeficiency virus infection of cells arrested in the cell cycle. Embo J 11(8):3053-8. Liberatore C, Capanni M, Albi N, Volpi I, Urbani E, Ruggeri L, Mencarelli A, Grignani F, Velardi A. 1999. Natural killer cell-mediated lysis of autologous cells modified by gene therapy. J Exp Med 189(12):1855-62. Loew R, Meyer Y, Kuehlcke K, Gama-Norton L, Wirth D, Hauser H, Stein S, Grez M, Thornhill S, Thrasher A and others. 2009. A new PG13-based packaging cell line for stable production of clinical-grade self-inactivating gamma-retroviral vectors using targeted integration.

In this context, the use of non-human cells would be strongly recommended, although the different glycosylation patterns of the envelope proteins could be an obstacle. For research purposes other human or monkey derived cells were tested (other 293 derived clones, HeLa, HT1080, TE671, COS1, COS-7, CV-1), although most of them showed reduced vector production titers. Yet, COS1 cells have shown to be capable of producing 3-4 times improved vector quality (expressed in infectious vector titer per ng of CA protein, p24), comparing with 293T cells (Smith and Shioda 2009).

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