Culture medium induced vimentin reorganization associates with enhanced baculovirus-mediated gene delivery

被引:16
|
作者
Mahonen, Anssi J. [1 ,2 ]
Makkonen, Kaisa-Emilia [2 ]
Laakkonen, Johanna P. [3 ]
Ihalainen, Teemu O. [3 ]
Kukkonen, Sari P. [1 ,2 ]
Kaikkonen, Minna U. [1 ,2 ]
Vihinen-Ranta, Maija [3 ]
Yla-Herttuala, Seppo [2 ,4 ,5 ,6 ]
Airenne, Kari J. [2 ]
机构
[1] Ark Therapeut Oy, FI-70210 Kuopio, Finland
[2] Univ Kuopio, AI Virtanen Inst, Dept Biotechnol & Mol Med, FIN-70211 Kuopio, Finland
[3] Univ Jyvaskyla, NanoSci Ctr, Dept Biol & Environm Sci, Jyvaskyla, Finland
[4] Kuopio Univ Hosp, SF-70210 Kuopio, Finland
[5] Univ Kuopio, Dept Med, SF-70210 Kuopio, Finland
[6] Univ Kuopio, Gene Therapy Unit, FIN-70211 Kuopio, Finland
基金
芬兰科学院;
关键词
Baculovirus; Intermediate filaments; Vimentin; Phosphorylation; Cell culture medium; Nuclear entry; INTERMEDIATE-FILAMENT NETWORK; NUCLEAR POLYHEDROSIS-VIRUS; MAMMALIAN-CELLS; IN-VIVO; RECOMBINANT BACULOVIRUS; HEPATOCELLULAR-CARCINOMA; HUMAN HEPATOCYTES; ESCHERICHIA-COLI; EXPRESSION; PHOSPHORYLATION;
D O I
10.1016/j.jbiotec.2009.11.003
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Baculoviruses can express transgenes under mammalian promoters in a wide range of vertebrate cells. However, the success of transgene expression is dependent on both the appropriate cell type and culture conditions. We studied the mechanism behind the substantial effect of the cell culture medium on efficiency of the baculovirus transduction in different cell lines. We tested six cell Culture mediums: the highest transduction efficiency was detected in the presence of RPMI 1640 medium. Vimentin, a major component of type III intermediate filaments, was reorganized in the optimized medium, which associated with enhanced nuclear entry of baculoviruses. Accordingly, the phosphorylation pattern of vimentin was changed in the studied cell lines. These results suggest that vimentin has an important role in baculovirus entry into vertebrate cells. Enhanced gene delivery in the optimized medium was observed also with adenoviruses and lentiviruses. The results highlight the general importance of the Culture medium in the assembly of the cytoskeleton network and in viral gene delivery. (C) 2009 Elsevier B.V. All rights reserved
引用
收藏
页码:111 / 119
页数:9
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