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Three-dimensional spheroid cultures of A549 and HepG2 cells exhibit different lipopolysaccharide (LPS) receptor expression and LPS-induced cytokine response compared with monolayer cultures

Jinsheng Xu; Liu, Jian; Abate, Wondwossen; Corry, David; Kaul, Baksho; Jackson, Simon K.

Authors

Jinsheng Xu

Jian Liu

Wondwossen Abate

David Corry

Baksho Kaul

Simon K. Jackson



Abstract

Lipopolysaccharide (LPS) is a potent modulator of pathogen-induced host inflammatory responses. Lipopolysaccharide signaling to host cells is correlated with the expression of well-characterized LPS receptors. We have developed three-dimensional (3-D) cell cultures (spheroids) that are more representative of in vivo conditions than traditional monolayer cultures and may provide novel in vitro models to study the inflammatory response. In this work, we have compared F-actin organization, LPS-induced pro-inflammatory cytokine response and LPS receptor expression between spheroid and monolayer cultures from A549 lung epithelial cells and HepG2 hepatocytes. Significant junctional F-actin was seen at the cell-cell contact points throughout the spheroids, while monolayer cells showed stress fibers of actin and more prominent F-actin localized at the cell base. A time course of cytokine release in response to LPS showed that A549 spheroids secreted persistently higher levels of interleukin (IL)-6 and IL-8 compared with monolayer cultures. Unlike monolayer cultures, HepG2 spheroids responded to LPS by releasing a significant level of IL-8. We identified a significant increase in the expression of CD14 and MD2 in these spheroids compared with monolayers, which may explain the enhanced cytokine response to LPS. Thus, we suggest that 3-D spheroid cell cultures are more typical of in vivo cell responses to LPS during the development of inflammation and would be a better in vitro model in inflammation studies. © SAGE Publications 2010.

Citation

Jinsheng Xu, , Liu, J., Abate, W., Jinsheng Xu, J., Corry, D., Kaul, B., & Jackson, S. K. (2011). Three-dimensional spheroid cultures of A549 and HepG2 cells exhibit different lipopolysaccharide (LPS) receptor expression and LPS-induced cytokine response compared with monolayer cultures. Innate Immunity, 17(3), 245-255. https://doi.org/10.1177/1753425910365733

Journal Article Type Article
Publication Date Jun 1, 2011
Journal Innate Immunity
Print ISSN 1753-4259
Electronic ISSN 1753-4267
Publisher SAGE Publications (UK and US)
Peer Reviewed Not Peer Reviewed
Volume 17
Issue 3
Pages 245-255
DOI https://doi.org/10.1177/1753425910365733
Keywords 3-D culture, LPS, LPS receptor, pro-inflammatory cytokine, F-actin
Public URL https://uwe-repository.worktribe.com/output/962040
Publisher URL http://dx.doi.org/10.1177/1753425910365733
Related Public URLs http://ini.sagepub.com/