title: Time-lapse ultrashort pulse microscopy of infection in three-dimensional versus two-dimensional culture environments reveals enhanced extra-chromosomal virus replication compartment formation.
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.
authors: Department of Biomedical Engineering, Texas A&M University, College Station, Texas, USA. holly.gibbs.bme@gmail.com
authors: Holly C
authors: HC
authors: Gibbs
authors: Garwin
authors: G
authors: Sing
authors: Juan Carlos
authors: JC
authors: González Armas
authors: Colin J
authors: CJ
authors: Campbell
authors: Peter
authors: P
authors: Ghazal
authors: Alvin T
authors: AT
authors: Yeh
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D003586:Cytomegalovirus Infections / Q000473:pathology / Q000821:virology*; D054884:Host-Pathogen Interactions / Q000502:physiology; D051379:Mice; D008853:Microscopy / Q000379:methods*; D041681:NIH 3T3 Cells; D059008:Time-Lapse Imaging / Q000379:methods*; D018406:Virus Physiological Phenomena; D014779:Virus Replication / Q000502:physiology; D014780:Viruses / Q000235:genetics