Involvement of hypoxia inducible factor activity in inevitable air-exposure treatment upon differentiation in a three-dimensional keratinocyte culture.
Involvement of hypoxia inducible factor activity in inevitable air-exposure treatment upon differentiation in a three-dimensional keratinocyte culture.
复制标题
三维角质形成细胞培养物分化时不可避免的空气暴露处理中缺氧诱导因子活性的参与。
DOI：
10.1111/febs.16707
复制
发表时间：
2023
期刊：
影响因子：
5.4
通讯作者：
and Hitomi K.
中科院分区：
文献类型：
--
作者：
Teshima H.;Endo M.;Furuyama Y.;Takama H.;Tsuji T.;Tatsukawa H.;Akiyama M.;and Hitomi K.
人皮肤表皮的形成可以通过三维（3D）角质形成细胞培养系统再现，其中在分化开始时空气暴露是不可避免的。在没有空气暴露的连续浸没培养中，即使使用分化相容的培养基，也没有诱导几种角质形成细胞特异性蛋白，导致异常表皮层的形成。为了阐明空气暴露促进角质形成细胞分化的机制，我们对浸没式和气-液相间培养系统之间的生物学特性进行了比较分析。通过转录组学分析，缺氧诱导因子（HIF）相关基因似乎在这些培养的细胞中发生了显着变化。在浸没培养中，HIF对其典型反应元件的转录活性增强，而空气暴露处理尽管HIF蛋白水平高，但转录活性显著降低。通过试剂和遗传操作调节HIF活性表明，减少但保留的HIF转录活性基本上参与了分化。此外，我们首次表明，在浸没培养系统中人工补充氧气可以恢复角质形成细胞分化，如在空气暴露培养中所观察到的那样。因此，我们在3D培养系统中机械地评估了HIF如何调节角质形成细胞的空气暴露依赖性分化。
Formation of the human skin epidermis can be reproduced by a three‐dimensional (3D) keratinocyte culture system, in which air‐exposure is inevitable upon initiation of differentiation. In the continuous submerged culture without air‐exposure, even with a differentiation‐compatible medium, several keratinocyte‐specific proteins were not induced resulting in the formation of aberrant epidermal layers. To clarify the mechanism by which air‐exposure promotes keratinocyte differentiation, we performed a comparative analysis on biological properties between submerged and air–liquid interphase culture systems. By transcriptomic analysis,hypoxia‐inducible factor (HIF)‐related genes appeared to significantly change in these cultured cells. In submerged culture, the transcriptional activity of HIF on its canonical response element was enhanced, while air‐exposure treatment drastically reduced the transcriptional activity despite the high HIF protein level. Regulating HIF activity through reagents and genetic manipulation revealed that the reduced but retained HIF‐transcriptional activity was essentially involved in differentiation. Furthermore, we showed, for the first time, that artificial supplementation of oxygen in the submerged culture system could restore keratinocyte differentiation as observed in the air‐exposed culture. Thus, we mechanistically evaluated how HIF regulates the air‐exposure‐dependent differentiation of keratinocytes in a 3D culture system.