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Specific substrates composed of collagen and fibronectin support the formation of epithelial cell sheets by MDCK cells lacking α-catenin or classical cadherins

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Abstract

The effect of the extracellular matrix substrates on the formation of epithelial cell sheets was studied using MDCK cells in which the α-catenin gene was disrupted. Although the mutant cells did not form an epithelial cell sheet in conventional cell culture, the cells formed an epithelial cell sheet when they were cultured on or in a collagen gel; the same results were not observed when cells were cultured on collagen-coated cover glasses or culture dishes. Moreover, the cells cultured on the cell culture inserts coated with fibronectin, Matrigel, or vitronectin formed epithelial cell sheets, whereas the cells cultured on cover glasses coated with these proteins did not form the structure, implying that the physical and chemical features of the substrates exert a profound effect on the formation of epithelial cell sheets. MDCK cells lacking the expression of E- and K-cadherins displayed similar properties. When the mutant MDCK cells were cultured in the presence of blebbistatin, they formed epithelial cell sheets, suggesting that myosin II was involved in the formation of these sheets. These cell sheets showed intimate cell–cell adhesion, and electron microscopy confirmed the formation of cell junctions. We propose that specific ECM substrates organize the formation of basic epithelial cell sheets, whereas classical cadherins stabilize cell–cell contacts and promote the formation of structures.

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Abbreviations

ECM:

Extracellular matrix

MDCK:

Madin-Darby canine kidney

TBS:

Tris-buffered saline

TPA:

12-O-tetradecanoylphorbol-13-acetate

DIC:

Differential interference contrast microscopy

SD:

Standard deviation

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Acknowledgements

We thank Mr. K. Kitayama and Mr. M. Yamaguchi for their technical assistance. We also thank Dr. K. Sekiguchi (Osaka University) for his valuable comments on this study. Human α-catenin cDNA was kindly provided by Dr. M. Ozawa (Kagoshima University).

Funding

This study was supported in part by Grants-in-Aid for Scientific Research from the Japan Society for Science Promotion (18K06230 to SH and 17K00981 to SO).

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Authors and Affiliations

  1. Department of Biology, School of Medicine, Kansai Medical University, 2-5-1 Shin-machi, Hirakata, Osaka, Japan

    Shintaro T. Suzuki & Shinji Hirano

  2. Department of Anatomical Sciences, School of Allied Health Sciences, Kitasato University, 1-15-1 Kitasato, Minami-ku, Sagamihara, Kanagawa, Japan

    Shuichi Obata

  3. Department of Histology and Cell Biology, School of Medicine, Yokohama City University, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa, Japan

    Shuichi Obata

  4. Department of Bioscience, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo, Japan

    Miwako Fujiwara

  5. Department of Microbiology, School of Medicine, Kansai Medical University, 2-5-1 Shin-machi, Hirakata, Osaka, Japan

    Jun-ichi Fujisawa

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  1. Shintaro T. Suzuki
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  2. Shuichi Obata
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Correspondence to Shintaro T. Suzuki.

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Suzuki, S.T., Obata, S., Fujiwara, M. et al. Specific substrates composed of collagen and fibronectin support the formation of epithelial cell sheets by MDCK cells lacking α-catenin or classical cadherins. Cell Tissue Res 385, 127–148 (2021). https://doi.org/10.1007/s00441-021-03448-1

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