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Subconfluent ARPE-19 Cells Display Mesenchymal Cell-State Characteristics and Behave like Fibroblasts, Rather Than
Preethi Golconda1, Mariana Andrade-Medina1, Adam Oberstein1
1Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicago, 835 South Wolcott Ave., Chicago, IL 60612, USA.
Abstract:
Human cytomegalovirus (HCMV) has a broad cellular tropism and epithelial cells are important physiological targets during infection. The retinal pigment epithelial cell line ARPE-19 has been used to model HCMV infection in epithelial cells for decades and remains a commonly used cell type for studying viral entry, replication, and the cellular response to infection. We previously found that ARPE-19 cells, despite being derived from an epithelial cell explant, express extremely low levels of canonical epithelial proteins, such as E-cadherin and EpCAM. Here, we perform comparative studies of ARPE-19 and additional epithelial cell lines with strong epithelial characteristics. We find that ARPE-19 cells cultured under subconfluent conditions resemble mesenchymal fibroblasts, rather than epithelial cells; this is consistent with previous studies showing that ARPE-19 cultures require extended periods of high confluency culture to maintain epithelial characteristics. By reanalyzing public gene expression data and using machine learning, we find evidence that ARPE-19 cultures maintained across many labs exhibit mesenchymal characteristics and that the majority of studies employing ARPE-19 use them in a mesenchymal state. Lastly, by performing experimental HCMV infections across mesenchymal and epithelial cell lines, we find that ARPE-19 cells behave like mesenchymal fibroblasts, producing logarithmic yields of cell-free infectious progeny, while cell lines with strong epithelial character exhibit an atypical infectious cycle and naturally restrict the production of cell-free progeny. Our work highlights important characteristics of the ARPE-19 cell line and suggests that subconfluent ARPE-19 cells may not be optimal for modeling epithelial infection with HCMV or other human viruses. It also suggests that HCMV biosynthesis and/or spread may occur quite differently in epithelial cells compared to mesenchymal cells. These differences could contribute to viral persistence or pathogenesis in epithelial tissues.
Insights
The ARPE-19 cell line, commonly used for human cytomegalovirus (HCMV) research, often exhibits mesenchymal, not epithelial, characteristics. This impacts how HCMV infection and spread are modeled in epithelial cells.
Area of Science:
- Virology
- Cell Biology
- Molecular Biology
Background:
- Human cytomegalovirus (HCMV) infects various cell types, including epithelial cells, which are crucial in disease.
- The ARPE-19 cell line is widely used to study HCMV in epithelial cells.
- ARPE-19 cells express low levels of epithelial markers and can exhibit mesenchymal characteristics.
Purpose of the Study:
- To compare ARPE-19 cells with other epithelial cell lines regarding their characteristics and response to HCMV infection.
- To investigate the cellular state (epithelial vs. mesenchymal) of ARPE-19 cells used in HCMV research.
- To determine if ARPE-19 cells are suitable models for epithelial HCMV infection.
Main Methods:
- Comparative analysis of ARPE-19 and other epithelial cell lines.
- Gene expression data reanalysis and machine learning.
- Experimental HCMV infections in mesenchymal and epithelial cell lines.
Main Results:
- ARPE-19 cells under standard culture conditions display mesenchymal, not epithelial, features.
- Public data and machine learning confirm widespread use of ARPE-19 in a mesenchymal state.
- ARPE-19 cells support logarithmic production of infectious HCMV, unlike epithelial cells that restrict cell-free progeny.
Conclusions:
- Subconfluent ARPE-19 cells may not accurately model epithelial HCMV infection.
- HCMV replication and spread may differ significantly between epithelial and mesenchymal cells.
- These cellular differences could influence HCMV persistence and pathogenesis in epithelial tissues.
