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. 2018 Mar 15;31(2):136–145. doi: 10.3967/bes2018.016

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

Ya Xiong CHEN a,^, Guang Cheng XIE b,^, Dong PAN a, Ya Rong DU a, Li Li PANG c, Jing Dong SONG c, Zhao Jun DUAN c, Bu Rong HU a,#
PMCID: PMC7134816  PMID: 29606192

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.

Key words: 3D cell culture, Human airway epithelium (HAE), Human rhinovirus C, Human bocavirus, Propagation

Biographies

Biographical notes of the first authors: CHEN Ya Xiong, male, born in 1984, MA, majoring in biochemistry and molecular biology

XIE Guang Cheng, male, born in 1982, PhD, majoring in pathogenic biology.

Footnotes

This work was supported by grants from the Major Project Specialized for Infectious Diseases of the Chinese Health and Family Planning Commission [2014ZX10004002-004-002, 2014ZX10004002-004-001]; Young Talent Scholar Plan of Higher School in Hebei Province [BJ2017008].

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