Introduction: Human Norovirus (HuNoV), a food-borne virus is the leading cause for acute gastroenteritis. However, its inability to propagate in vitropersists as major challenge in understanding HuNoV biology.Objective: This study aims to determine an effective culture system for HuNoV.Methods: The Caco-2 cells were cocultured with Raji B cells on alginate hydrogel beads. Scanning electron microscopy (SEM) was performed to confirmthe three-dimensional (3D) cells morphology. Western blot (WB) analysis was performed to detect protein markers expressed by Microfold (M) cells.Results: Optimization of Caco-2 cells monoculture in the alginate hydrogel beads showed optimum number of cells of 1 × 106 cells/ml, indicatedby the intact structure of the beads. Result of SEM showed clear structure of monoculture in the alginate hydrogel beads indicated by the presenceof smooth and regular apical surface while the coculture showed reduced apical surface of M cells. The result of WB showed downregulation ofUlex europaeus antibody expression.Conclusion: It is evident that the expression of M cells grown in 3D alginate hydrogel beads was successful, indicated by the structural morphologyseen under SEM as well as expression of protein marker by M cells. This established in vitro system is highly potential for cultivation of HuNoV.
Human noroviruses (HuNoV) are recognized as the primary cause for non-bacterial acute gastroenteritis. The HuNoV outbreak was firstly identified at Norwalk, Ohio and formerly known as 'winter vomiting disease' due to the illness frequently strikes during winter season. Infection of HuNoV is estimated to affects all age groups in both developed and developing country, by presenting symptoms such as nausea, vomiting, diarrhea and low-grade fever. There are approximately seven genogroups of noroviruses have been identified in which genogroup II.4 is the dominant HuNoV causing disease to human. Despite their significant impact on the economic and health burden, noroviruses remain to be poorly characterized RNA virus due to the lack of reproducible in vitro culture system for the HuNoV propagation, thus hampering understanding of its pathogenesis. Several approaches including the cultivation trials and the discovery of murine Norovirus (MuNoV) as the model system have expanded our knowledge in Norovirus biology and life cycle. However, to date there are no reliable culture system for HuNoV has been reported. The development of 3D co-culture system has shown a promising future as the improvised system that can be utilized not only for HuNoV cultivation, but also to other viruses. In this review, various attempts to cultivate the HuNoV are discussed to emphasise the future strategy in obtaining reproducible culture system. This review aims to find the gaps in the search of a reliable and reproducible propagation system thus enhancing the understanding on HuNoV biology and pathogenesis for future development of accurate detection method and vaccine production.
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