3D Tissue Culture Model for Virology Studies

Abstract

Infectious diseases, including recent outbreaks of H7N9 influenza, Ebola, Zika, and SARS-CoV-2, remain significant global health threats. While traditional two-dimensional (2D) cell cultures have long been the cornerstone of virology research, their inability to replicate complex in vivo microenvironments, such as cell-cell interactions, apical-basal polarity, and extracellular signaling gradients, limits their utility for studying viral pathogenesis and drug responses. Three-dimensional (3D) culture systems overcome these limitations by providing physiologically relevant platforms that better mimic native tissue environments. Among the commonly used methods, the hanging-drop method enables spheroid formation by suspending cell droplets, allowing natural cell aggregation at the liquid-air interface. The pHEMA method creates a nonadhesive surface through a poly-2-hydroxyethyl methacrylate coating, ensuring uniform spheroid sizes and minimal cell-matrix interactions. The Matrigel embedding method embeds cells in a growth factor-reduced extracellular matrix, supporting cell-matrix interactions, tissue morphogenesis, and differentiation. Finally, the inlet-well-based hanging-drop method employs a specialized inlet-well system, combining hanging-drop formation with controlled transfer to receiving wells while maintaining humidity and minimizing evaporation. These versatile methods facilitate studies on viral replication, infectivity, and antiviral screening, offering reproducible, high-fidelity models for understanding host-virus interactions and advancing therapeutic development.

Keywords: 3D cell culture; Hanging-drop systems; Matrigel; Spheroid; pHEMA.