Three-dimensional Culture of Human Airway Epithelium in Matrigel for Evaluation of Human Rhinovirus C and Bocavirus Infections
| Autor: | CHEN, Ya Xiong, XIE, Guang Cheng, PAN, Dong, DU, Ya Rong, PANG, Li Li, SONG, Jing Dong, DUAN, Zhao Jun, HU, Bu Rong |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
3D cell culture
Human rhinovirus C Virus Cultivation Primary Cell Culture Human airway epithelium (HAE) Enzyme-Linked Immunosorbent Assay Epithelial Cells Respiratory Mucosa Real-Time Polymerase Chain Reaction Article Parvoviridae Infections Drug Combinations Human bocavirus Enterovirus Infections Humans Proteoglycans Collagen Laminin Propagation Enterovirus |
| Zdroj: | Biomedical and Environmental Sciences |
| ISSN: | 2214-0190 0895-3988 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|