Table 5.

In vitro cell models of viral infection based on bioreactor-perfused cultures

Refs.	Pathogen	Cell type/s	Cell culture configuration
67-68	SARS-CoV.	Primary human bronchial-tracheal cells (HBTCs) used to establish a human lung tissue-like assembly (TLA) with an overlay of human bronchial epithelial (BEAS-2B) cells. 2D control established with BEAS-2B monoculture.	3D tissue dynamic culture system established using a Rotating Wall Vessel (RWV) bioreactor. Static 2D controls: tissue culture flasks of near confluent BEAS-2B monocultures.
70	Hepatitis B virus (HBV).	Primary Human Hepatocytes (PHH) and co-culture with primary Kupffer cells (KC).	Perfused 3D bioreactor. Medium was recirculated via a pneumatically driven micro-pump, and collagen-coated scaffolds were used for cell adherence. Each plate consisted of 12 individual bioreactors, in which the flow rate was regulated. 3D perfused: PHH seeded in 3D scaffolds. Non-perfused controls: 3D spheroid, static 2D PHH, and self-assembling co-cultures (SACC) of PHH.
68-69	Varicella zoster virus (VZV).	Normal human neural progenitor (NHNP) cells used to establish human neuronal tissue- like assembly (TLA). Gene mutations, infection study and VZV genome maintenance were performed on the TLA and validated with a 2D control of human melanoma (MeWo) cells.	Optimized 3D tissue culture systems created using a Rotating Wall Vessel (RWV) bioreactor. The 3D model was compared to a 2D control of NHNP cells.
71	Adenovirus carrying enhanced green fluorescent protein (AdEGFP).	Mouse embryonic fibroblasts (MEFs) and human foreskin fibroblasts (HFFs) both for microfluidic study and static control.	Microfluidic device composed of a supporting glass slide with a PDMS slab carved to accommodate the cell culture coverslip, a membrane-based vacuum system for the reversible sealing, and microfluidic channels delivering fluids to the cultured cells. The control was performed in static conditions.
106	Enhanced green fluorescent protein (EGFP) plasmid.	3T3 fibroblasts used for 2D control, static and dynamic conditions.	Microfluidic perfused device for dynamic cell culture and virus infection in a micro channel, control was performed in 2D conventional culture, and in the microfluidic device was evaluated in both static and dynamic conditions.