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Vimentin is necessary for colony growth of human diploid keratinocytes

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Abstract

The role of vimentin (Vim) in diploid epithelial cells is not well known. To understand its biological function, we cultured human epidermal keratinocytes under conditions that support migration, proliferation, stratification and terminal differentiation. We identified a keratinocyte subpopulation that shows a p63+/α5β1bright phenotype and displays Vim intermediate filaments (IFs) besides their keratin IF network. These cells were mainly located at the proliferative/migratory rim of the growing colonies; but also, they were scarce and scattered or formed small groups of basal cells in confluent stratified epithelia. Stimulation of cells with EGF and wounding experiments in confluent arrested epithelia increased the number of Vim+ keratinocytes in an extent higher to the expected for a cell population doubling. BrdU labeling demonstrated that most of the proliferative cells located at the migratory border of the colony have Vim, in contrast with proliferative cells located at the basal layer at the center of big colonies which lacked of Vim IFs, suggesting that Vim expression was not solely linked to proliferation. Therefore, we silenced Vim mRNA in the cultured keratinocytes and observed an inhibition of colony growth. Such results, together with long-term cultivation assays which showed that Vim might be associated to pattern formation in cultured epithelia, suggest that Vim expression is essential for a highly motile phenotype, which is necessary for keratinocyte colony growth and possibly for development and wound healing. Vim+/p63+/α5β1bright epithelial cells may play a significant physiological role in embryonic morphogenetic movements; wound healing and other pathologies such as carcinomas and hyperproliferative diseases.

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Acknowledgments

This work was supported in part through grant PICDS08-8 from Instituto de Ciencia y Tecnología del Distrito Federal (ICyTDF) and Consejo Nacional de Ciencia y Tecnología (CONACyT) grants 1314P-N9507 and G28272-N. The authors would like to thank Mrs. Erika Sánchez Guzmán for technical assistance. The authors would also like to thank Ms. Maria Elena Rojano for her administrative assistance and Mr. Alberto Rodriguez, Mr. Crescencio Flores, and Mrs. Columba Guadarrama for technical assistance in preparing all of the materials.

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  1. Department of Cell Biology, Centro de Investigación y de Estudios Avanzados del IPN Apdo, Postal 14-740, 07000, Mexico City, Mexico

    Federico Castro-Muñozledo, Cristina Velez-DelValle, Meytha Marsch-Moreno, Miriam Hernández-Quintero & Walid Kuri-Harcuch

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  1. Federico Castro-Muñozledo
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  2. Cristina Velez-DelValle
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  3. Meytha Marsch-Moreno
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  4. Miriam Hernández-Quintero
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Correspondence to Federico Castro-Muñozledo.

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Federico Castro-Muñozledo and Cristina Velez-DelValle have contributed equally to this work.

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Castro-Muñozledo, F., Velez-DelValle, C., Marsch-Moreno, M. et al. Vimentin is necessary for colony growth of human diploid keratinocytes. Histochem Cell Biol 143, 45–57 (2015). https://doi.org/10.1007/s00418-014-1262-6

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