Three-dimensional Culture of Human Airway Epithelium in Matrigel for Evaluation of Human Rhinovirus C and Bocavirus Infections.

Ya Xiong Chen; Guang Cheng Xie; Dong Pan; Ya Rong DU; Li Li Pang; Jing Dong Song; Zhao Jun Duan; Bu Rong HU

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Three-dimensional Culture of Human Airway Epithelium in Matrigel for Evaluation of Human Rhinovirus C and Bocavirus Infections.

Author: Ya Xiong CHEN ¹ ; Guang Cheng XIE ² ; Dong PAN ¹ ; Ya Rong DU ¹ ; Li Li PANG ³ ; Jing Dong SONG ³ ; Zhao Jun DUAN ³ ; Bu Rong HU ¹
Author Information

1. Key Laboratory of Heavy Ion Radiation Biology and Medicine of Chinese Academy of Sciences & Key Laboratory of Space Radiobiology of Gansu Province, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou 730000, Gansu, China.
2. Department of Pathogenic Biology, Chengde Medical University, Chengde 067000, Hebei, China.
3. Department for Viral Diarrhea, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Publication Type:Journal Article
Keywords: 3D cell culture; Human airway epithelium (HAE); Human bocavirus; Human rhinovirus C; Propagation
MeSH: Collagen; Drug Combinations; Enterovirus; growth & development; isolation & purification; Enterovirus Infections; virology; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; virology; Human bocavirus; growth & development; isolation & purification; Humans; Laminin; Parvoviridae Infections; virology; Primary Cell Culture; methods; Proteoglycans; Real-Time Polymerase Chain Reaction; Respiratory Mucosa; virology; Virus Cultivation
From: Biomedical and Environmental Sciences 2018;31(2):136-145
CountryChina
Language:English
Abstract: 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.