Cell jamming, stratification and p63 expression in cultivated human corneal epithelial cell sheets
Cell jamming, stratification and p63 expression in cultivated human corneal epithelial cell sheets
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DOI：
10.1038/s41598-020-64394-6
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发表时间：
2020-06-09
影响因子：
4.6
通讯作者：
Nishida, Kohji
中科院分区：
文献类型：
--
作者：
Baba, Koichi;Sasaki, Kei;Nishida, Kohji
利用培养的人角膜上皮细胞片进行角膜缘上皮干细胞移植已成功地用于治疗角膜缘干细胞缺乏症。在这里，我们报道了一项对培养的人角膜上皮细胞片质量的调查，使用了14天内每20分钟一次的细胞培养过程的时间推移成像，以确定细胞堵塞的程度，在这种现象中，细胞变得更小，更圆，更不活跃的扩张。同时，我们还评估了重要的角膜上皮干细胞标记物p63的表达。细胞堵塞的发生是可变的和短暂的，但总是与细胞膜的增厚和分层有关。在培养的前9天，P63在所有扩张的细胞膜中都存在，但它的存在并不总是与细胞膜的分层相关。P63的表达也不一定在分层的细胞膜中持续存在。因此，对细胞堵塞的评估可以为培养的人角膜上皮细胞片的质量和再生潜力提供重要的线索。
Corneal limbal epithelial stem cell transplantation using cultivated human corneal epithelial cell sheets has been used successfully to treat limbal stem cell deficiencies. Here we report an investigation into the quality of cultivated human corneal epithelial cell sheets using time-lapse imaging of the cell culture process every 20minutes over 14 days to ascertain the level of cell jamming, a phenomenon in which cells become smaller, more rounded and less actively expansive. In parallel, we also assessed the expression of p63, an important corneal epithelial stem cell marker. The occurrence of cell jamming was variable and transient, but was invariably associated with a thickening and stratification of the cell sheet. p63 was present in all expanding cell sheets in the first 9 days of culture, but it's presence did not always correlate with stratification of the cell sheet. Nor did p63 expression necessarily persist in stratified cell sheets. An assessment of cell jamming, therefore, can shed significant light on the quality and regenerative potential of cultivated human corneal epithelial cell sheets.