Graduate School of Pharmaceutical Sciences, Nagoya University, Nagoya, Japan.
Graduate School of Biostudies, Kyoto University, Kyoto, Japan.
Biosci Biotechnol Biochem. 2024 Jul 22;88(8):932-940. doi: 10.1093/bbb/zbae070.
A 3-dimensional culture system of keratinocytes achieves cornification as a terminal differentiation that can mimic the formation of stratified epidermis. At the onset of keratinocyte differentiation, air-exposure treatment is essential for promotion. We have previously reported that the stimulation of differentiation is accompanied by downregulation of the transcriptional activity of the hypoxia-inducible factor (HIF) and also found that rocking treatment of cultured keratinocytes in the submerged condition restored their differentiation. A comparative study of cultured keratinocytes with and without rocking was then carried out to investigate the characteristics of the recovered differentiation by morphological and biochemical analyses. In addition, transcriptome analysis revealed the expected similar pattern between air-exposed and rocking cultures, including HIF-regulating transcripts. Furthermore, the promotive effect of rocking treatment was impaired under hypoxic culture conditions (1% O2). We showed that the restored promotion of differentiation by rocking culture is mainly due to the abrogation of transcriptional events by hypoxia.
三维培养的角质形成细胞可实现角化这一终末分化，可模拟分层表皮的形成。在角质形成细胞分化开始时，空气暴露处理对促进分化至关重要。我们之前曾报道过，分化的刺激伴随着缺氧诱导因子 (HIF) 的转录活性下调，并且还发现，在淹没条件下摇动培养的角质形成细胞可以恢复其分化。然后进行了有和没有摇动的培养角质形成细胞的比较研究，通过形态和生化分析来研究恢复分化的特征。此外，转录组分析显示，空气暴露和摇动培养之间存在预期的相似模式，包括 HIF 调节转录本。此外，在低氧培养条件（1%O2）下，摇动处理的促进作用受损。我们表明，摇动培养恢复分化的促进作用主要归因于缺氧引起的转录事件的中止。
