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Biomedical and Environmental Sciences ; (12): 136-145, 2018.
Article Dans Anglais | WPRIM | ID: wpr-776071

Résumé

OBJECTIVE@#Newly identified human rhinovirus C (HRV-C) and human bocavirus (HBoV) cannot propagate in vitro in traditional cell culture models; thus obtaining knowledge about these viruses and developing related vaccines are difficult. Therefore, it is necessary to develop a novel platform for the propagation of these types of viruses.@*METHODS@#A platform for culturing human airway epithelia in a three-dimensional (3D) pattern using Matrigel as scaffold was developed. The features of 3D culture were identified by immunochemical staining and transmission electron microscopy. Nucleic acid levels of HRV-C and HBoV in 3D cells at designated time points were quantitated by real-time polymerase chain reaction (PCR). Levels of cytokines, whose secretion was induced by the viruses, were measured by ELISA.@*RESULTS@#Properties of bronchial-like tissues, such as the expression of biomarkers CK5, ZO-1, and PCK, and the development of cilium-like protuberances indicative of the human respiration tract, were observed in 3D-cultured human airway epithelial (HAE) cultures, but not in monolayer-cultured cells. Nucleic acid levels of HRV-C and HBoV and levels of virus-induced cytokines were also measured using the 3D culture system.@*CONCLUSION@#Our data provide a preliminary indication that the 3D culture model of primary epithelia using a Matrigel scaffold in vitro can be used to propagate HRV-C and HBoV.


Sujets)
Humains , Collagène , Association médicamenteuse , Enterovirus , Infections à entérovirus , Virologie , Test ELISA , Cellules épithéliales , Virologie , Bocavirus humain , Laminine , Infections à Parvoviridae , Virologie , Culture de cellules primaires , Méthodes , Protéoglycanes , Réaction de polymérisation en chaine en temps réel , Muqueuse respiratoire , Virologie , Culture virale
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