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Propagation of viruses on micropatterned host cells
Author(s) -
Endler Elizabeth E.,
Duca Karen A.,
Nealey Paul F.,
Whitesides George M.,
Yin John
Publication year - 2003
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10516
Subject(s) - vesicular stomatitis virus , baby hamster kidney cell , microbiology and biotechnology , cell culture , virus , cytoskeleton , cell , biology , biophysics , in vitro , cell fusion , viral entry , virology , chemistry , viral replication , biochemistry , genetics
We have developed a technique to characterize the in vitro propagation of viruses. Microcontact printing was used to generate linear arrays of alkanethiols on gold surfaces, which served as substrates for the patterned culture of baby hamster kidney (BHK‐21) cells. Vesicular stomatitis virus (VSV) was added to unpatterned cell reservoirs adjacent to the patterned cells and incubated, setting in motion a continuously advancing viral infection into the patterned cells. At different incubation times, multiple arrays were chemically fixed to stop the viral propagation. Viral propagation distances into the patterned cells were determined by indirect immunofluorescent labeling and visualization of the VSV surface glycoprotein (G). The infection spread at approximately 50 μm/h in the 140‐μm lines. Moreover, different temporal stages of the infection process were simultaneously visualized along individual lines. These stages included initiation of infection, based on G protein expression; cell–cell fusion, based on virus‐induced clustering of cell nuclei; and cytoskeletal degradation, based on localized release of cells from the surface. This work sets a foundation for parallel, high‐throughput characterization of viral and cellular processes. © 2003 Wiley Periodicals Inc. Biotechnol Bioeng 81: 719–725, 2003.