A novel tool for the unbiased characterization of epithelial monolayer development in culture
The function of an epithelial tissue is intertwined with its architecture. Epithelial tissues are often described as pseudo-two-dimensional, but this view may be partly attributed to experimental bias: many model epithelia, including cultured cell lines, are easiest to image from the "top-down....
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Published in | Molecular biology of the cell Vol. 34; no. 4; p. ar25 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
The American Society for Cell Biology
01.04.2023
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Subjects | |
Online Access | Get full text |
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Summary: | The function of an epithelial tissue is intertwined with its architecture. Epithelial tissues are often described as pseudo-two-dimensional, but this view may be partly attributed to experimental bias: many model epithelia, including cultured cell lines, are easiest to image from the "top-down." We measured the three-dimensional architecture of epithelial cells in culture and found that it varies dramatically across cultured regions, presenting a challenge for reproducibility and cross-study comparisons. We therefore developed a novel tool (Automated Layer Analysis, "ALAn") to characterize architecture in an unbiased manner. Using ALAn, we find that cultured epithelial cells can organize into four distinct architectures and that architecture correlates with cell density. Cells exhibit distinct biological properties in each architecture. Organization in the apical-basal axis is determined early in monolayer development by substrate availability, while disorganization in the apical-basal axis arises from an inability to form substrate connections. Our work highlights the need to carefully control for three-dimensional architecture when using cell culture as a model system for epithelial cell biology and introduces a novel tool, built on a set of rules that can be widely applied to epithelial cell culture. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 These authors should be considered co–first authors. Author contributions: D.T.B. and C.C. conceived the project. D.T.B., C.C., and N.S.D. designed ALAn. C.C., N.S.D., M.V. and J.A.G. coded ALAn. N.S.D., C.C., Q.J., A.S., and J.A.G. performed cultured cell experiments and analysis. D.T.B., C.C., N.S.D., and T.M.F. made figures and wrote the paper. Conflict of interest: The authors declare no competing interests. |
ISSN: | 1059-1524 1939-4586 1939-4586 |
DOI: | 10.1091/mbc.E22-04-0121 |