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Journal of the Association for Research in Otolaryngology

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05.04.2018 | Research Article

Transcription and microRNA Profiling of Cultured Human Tympanic Membrane Epidermal Keratinocytes

verfasst von: Peder Aabel, Tor Paaske Utheim, Ole Kristoffer Olstad, Helge Rask-Andersen, Rodney James Dilley, Magnus von Unge

Erschienen in: Journal of the Association for Research in Otolaryngology | Ausgabe 3/2018

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Abstract

The human tympanic membrane (TM) has a thin outer epidermal layer which plays an important role in TM homeostasis and ear health. The specialised cells of the TM epidermis have a different physiology compared to normal skin epidermal keratinocytes, displaying a dynamic and constitutive migration that maintains a clear TM surface and assists in regeneration. Here, we characterise and compare molecular phenotypes in keratinocyte cultures from TM and normal skin. TM keratinocytes were isolated by enzymatic digestion and cultured in vitro. We compared global mRNA and microRNA expression of the cultured cells with that of human epidermal keratinocyte cultures. Genes with either relatively higher or lower expression were analysed further using the biostatistical tools g:Profiler and Ingenuity Pathway Analysis. Approximately 500 genes were found differentially expressed. Gene ontology enrichment and Ingenuity analyses identified cellular migration and closely related biological processes to be the most significant functions of the genes highly expressed in the TM keratinocytes. The genes of low expression showed a marked difference in homeobox (HOX) genes of clusters A and C, giving the TM keratinocytes a strikingly low HOX gene expression profile. An in vitro scratch wound assay showed a more individualised cell movement in cells from the tympanic membrane than normal epidermal keratinocytes. We identified 10 microRNAs with differential expression, several of which can also be linked to regulation of cell migration and expression of HOX genes. Our data provides clues to understanding the specific physiological properties of TM keratinocytes, including candidate genes for constitutive migration, and may thus help focus further research.

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Titel: Transcription and microRNA Profiling of Cultured Human Tympanic Membrane Epidermal Keratinocytes
verfasst von: Peder Aabel
Tor Paaske Utheim
Ole Kristoffer Olstad
Helge Rask-Andersen
Rodney James Dilley
Magnus von Unge
Publikationsdatum: 05.04.2018
Verlag: Springer US
Erschienen in: Journal of the Association for Research in Otolaryngology / Ausgabe 3/2018
Print ISSN: 1525-3961
Elektronische ISSN: 1438-7573
DOI: https://doi.org/10.1007/s10162-018-0660-1

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Transcription and microRNA Profiling of Cultured Human Tympanic Membrane Epidermal Keratinocytes

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