Culture medium induced vimentin reorganization associates with enhanced baculovirus-mediated gene delivery.
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.
Mähönen
Anssi J
AJ
A.I. Virtanen Institute, Department of Biotechnology and Molecular Medicine, University of Kuopio Kuopio, Finland. anssi.mahonen@arktherapeutics.com
Makkonen
Kaisa-Emilia
KE
Laakkonen
Johanna P
JP
Ihalainen
Teemu O
TO
Kukkonen
Sari P
SP
Kaikkonen
Minna U
MU
Vihinen-Ranta
Maija
M
Ylä-Herttuala
Seppo
S
Airenne
Kari J
KJ
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism*; D004279:DNA, Viral / Q000235:genetics*; D006801:Humans; D014161:Transduction, Genetic / Q000379:methods*; D014746:Vimentin / Q000737:chemistry / Q000378:metabolism*
D003470:Culture Media; D004279:DNA, Viral; D014746:Vimentin