Propagation of viruses on micropatterned host cells.
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 microm/h in the 140-microm 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.
Endler
Elizabeth E
EE
Department of Chemical Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, Wisconsin 53706-1691, USA.
Duca
Karen A
KA
Nealey
Paul F
PF
Whitesides
George M
GM
Yin
John
J
D000818:Animals; D000831:Animals, Newborn; D002448:Cell Adhesion / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D018914:Cells, Immobilized / Q000166:cytology / Q000502:physiology; D006224:Cricetinae; D007668:Kidney / Q000166:cytology* / Q000502:physiology / Q000821:virology*; D008562:Membrane Glycoproteins / Q000378:metabolism; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D015336:Molecular Probe Techniques; D009038:Motion; D018353:Rhabdoviridae Infections / Q000473:pathology* / Q000503:physiopathology; D014721:Vesicular stomatitis Indiana virus / Q000254:growth & development / Q000302:isolation & purification / Q000502:physiology* / Q000648:ultrastructure*; D014759:Viral Envelope Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism; D014776:Virus Cultivation / Q000379:methods
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D008567:Membranes, Artificial; D014759:Viral Envelope Proteins; D014764:Viral Proteins