Vector producer cells

Gene Therapies. The types of vectors that have been used or proposed for gene transduction include retrovirus, adenovirus, adeno-associated viruses, other viruses (e.g., herpes, vaccinia, etc.), and plasmid DNA. Methods for gene introduction include ex vivo replacement, drug delivery, marker studies, and others and in vivo, viral vectors, plasmid vectors, and vector producer cells. 

Vector Producer Cells. The direct administration of retroviral vector producer cells (e.g., murine cells producing HTK vector) to patients. 

Culver, K.W., Ram, Z., Wallbridge, S., Ishii, H., Oldfield, E.H., and Blaese, R.M. (1992). In vivo gene transfer with retroviral vector-producer cells for treatment of experimental brain tumors. Science 256,1550 1552. 

Z. Ram, K.W. Culver, E.M. Oshiro, J.J. Viola, H.L. DeVroom, E. Otto, Therapy of malignant brain tumors by intratumoral implantation of retroviral vector-producing cells. Nature Med. 3 (1997) 1354-1361. 

The desired gene/cDNA is normally amplified, sequenced and then introduced into an expression vector that facilitates its introduction and expression (transcription and translation) in an appropriate producer cell type. All recombinant therapeutic proteins approved to date are produced in E. coli, S. cerevisiae or in animal cell lines (mainly CHO or BHK cells). The general... 

Loiler, S., DiFronzo, N. and Holland, C. (1997) Gene transfer to human cells using retrovirus vectors produced hy a new polytropic packaging cell line. J VirolTl, 4825-4828. 

Other protocols involve cell lines with integrated rep/cap cassettes (Clark et al., 1995 Gao et al., 1998 Liu et al., 1999 Chadeuf et al., 2000 Mathews et al., 2002 Qiao et al., 2002a,b) infected with adenovirus or, alternatively, a recombinant herpesvirus system has been used to provide both helper virus function and rep/cap (Conway et al., 1997, 1999). In a switch away from using mammalian cell and helper virus production systems, rAAV vectors have been made in insect cells where the AAV genes are expressed under the control of insect promoters and the traditional helper virus gene products are not required (Urabe et al., 2002). Stable producer cell... 

Mathews, L. C., Gray, J. T., Gallagher, M. R. and Snyder, R. O. (2002). Recombinant adeno-associated viral vector production using stable packaging and producer cell lines. Methods Enzymol. 346, 393-413. 

PER.C6 cells have been used extensively for the production of gene therapy vectors. This cell line was derived from the immortalization of human embryonic retina cells through the use of adenovirus El gene (Fallaux et al., 1998). This cell line has been well characterized since its establishment, and no retroviruses or adventitious viruses have been detected in it. This cell line is easily adapted to different growth conditions and stably produces high levels of recombinant proteins. 

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