[Establishment of an Epstein-Barr virus infection model using human nasal organoids].

OBJECTIVES: To develop an cost-effective and convenient method for culturing human nasal organoids to establish an in vitro Epstein-Barr virus (EBV) infection model. METHODS: Nasal polyp tissue obtained from surgery was routinely washed, cut, digested and filtered to obtain cell clusters. The cell clusters were then expanded into undifferentiated nasal organoids through dynamic suspension culture under matrix-free conditions, followed by a 14-day differentiation induction treatment to obtain differentiated nasal organoids. The major cellular components of the organoids were identified by immunofluorescence staining and immunohistochemistry. The nasal organoids were infected by EBV in vitro, and viral replication was verified by detecting the expressions of the virus-specific genes EBNA1 and BALF5 using RT-qPCR and immunofluorescence staining. RESULTS: Nasal organoids consisting of basal cells, mucous cells, and ciliated cells in a martigel-free system were obtained successfully by dynamic suspension culture. The differentiated nasal organoids expressed high levels of EBV-associated receptors EphA2, NRP1, and NMHCII-A. Both the undifferentiated and differentiated nasal organoids could be infected by EBV. Viral replication in the organoids increased with the viral exposure load, and the differentiated organoids appeared more permissive to viral replication. CONCLUSIONS: The matrigel-free dynamic suspension culture method is economical and simple for constructing nasal organoids, which can be used as a model of EBV infection for studies of epithelial EBV infection.