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- Title
Time-lapse ultrashort pulse microscopy of infection in three-dimensional versus two-dimensional culture environments reveals enhanced extra-chromosomal virus replication compartment formation.
- Authors
Gibbs, Holly C.; Sing, Garwin; González Armas, Juan Carlos; Campbell, Colin J.; Ghazal, Peter; Yeha, Alvin T.
- Abstract
The mechanisms that enable viruses to harness cellular machinery for their own survival are primarily studied in cell lines cultured in two-dimensional (2-D) environments. However, there are increasing reports of biological differences between cells cultured in 2-D versus three-dimensional (3-D) environments. Here we report differences in host-virus interactions based on differences in culture environment. Using ultrashort pulse microscopy (UPM), a form of two-photon microscopy that utilizes sub-10-fs pulses to efficiently excite fluorophores, we have shown that de novo development of extra-chromosomal virus replication compartments (VRCs) upon murine cytomegalovirus (mCMV) infection is markedly enhanced when host cells are cultured in 3-D collagen gels versus 2-D monolayers. In addition, time-lapse imaging revealed that mCMV-induced VRCs have the capacity to grow by coalescence. This work supports the future potential of 3-D culture as a useful bridge between traditional monolayer cultures and animal models to study host-virus interactions in a more physiologically relevant environment for the development of effective anti-viral therapeutics. These advances will require broader adoption of modalities, such as UPM, to image deep within scattering tissues.
- Subjects
HOST-virus relationships; CELL lines; MICROSCOPY; FLUOROPHORES; CYTOMEGALOVIRUS diseases; CELL culture
- Publication
Journal of Biomedical Optics, 2013, Vol 18, Issue 3, p1
- ISSN
1083-3668
- Publication type
Article
- DOI
10.1117/1.JBO.18.3.031111