Immobilization of 293 cells using porous support particles for adenovirus vector production |
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Authors: | Naoya Morishita Tomohisa Katsuda Shuji Kubo Akinobu Gotoh Hideki Yamaji |
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Affiliation: | (1) Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1–1 Rokkodai, Nada, Kobe 657–8501, Japan;(2) Institute for Advanced Medical Sciences, Hyogo College of Medicine, 1–1 Mukogawa, Nishinomiya 663–8501, Japan; |
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Abstract: | Adenovirus vector production by anchorage-independent 293 cells immobilized using porous biomass support particles (BSPs) was investigated in static and shake-flask cultures for efficient large-scale production of adenovirus vectors for gene therapy applications. The density of cells immobilized within BSPs was evaluated by measuring their WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt) reduction activity. In shake-flask culture, 293-F cells, which were adapted to serum-free suspension culture, were not successfully retained within reticulated polyvinyl formal (PVF) resin BSPs (2 × 2 × 2 mm cubes) with matrices of relatively small pores (pore diameter 60 μm). When the BSPs were coated with a cationic polymer polyethyleneimine, a high cell density of more than 107 cells cm−3-BSP was achieved in both static and shake-flask cultures with regular replacement of the culture medium. After infection with an adenovirus vector carrying the enhanced green fluorescent protein gene (Ad EGFP), the specific Ad EGFP productivity of the immobilized cells was comparable to the maximal productivity of non-immobilized 293-F cells by maintaining favorable conditions in the culture environment. |
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Keywords: | Adenovirus vector 293 cell Viral vector production Immobilization Biomass support particles Polyvinyl formal resin Polyethyleneimine Gene therapy |
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