Investigation of K14/K5 as a stem cell marker in the limbal region of the bovine cornea.
Investigation of K14/K5 as a stem cell marker in the limbal region of the bovine cornea.
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DOI：
10.1371/journal.pone.0013192
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发表时间：
2010-10-06
期刊：
影响因子：
3.7
通讯作者：
Connon CJ
中科院分区：
文献类型：
--
作者：
Chen B;Mi S;Wright B;Connon CJ
通过特定的分子标记从角膜上皮细胞群中鉴定干细胞已经进行了研究。P63、ABCG2和K14/K5的表达都与哺乳动物角膜上皮干细胞有关。在这里，我们报道了K14/K5作为角膜缘干细胞标志物的局限性。采用免疫组织化学、Western blotting和Real time PCR检测K14/K5在牛角膜缘和角膜中央上皮细胞中的表达，并进行比较。一项功能研究也包括了K5/14在培养的角膜缘上皮细胞中进行强制分化的表达变化。在角膜缘区和中央角膜的静止上皮细胞中均检测到K14（或其伴侣K5）的表达。K14主要定位于角膜边缘的基底上皮细胞和中央角膜的基底上上皮细胞。Western blot结果显示，角膜缘和角膜中央均有K14表达（角膜缘较高）。同样，定量实时PCR发现K5， K14的伴侣，在角膜中央和角膜缘都有表达。在培养中强制分化后，角膜缘上皮细胞显示出K5/14基因/蛋白表达水平的增加，与已知分化标志物的可预测上升相一致。K14和它的伙伴K5不仅局限于角膜缘，而且也局限于牛角膜中央上皮细胞，这表明K14/K5不是角膜缘特异性的。此外，在强迫分化的角膜缘上皮细胞培养中，K14/K5的表达水平没有降低（实际上是增加），这表明K14/K5是体外未分化细胞的不可靠的制造物。
Identification of stem cells from a corneal epithelial cell population by specific molecular markers has been investigated previously. Expressions of P63, ABCG2 and K14/K5 have all been linked to mammalian corneal epithelial stem cells. Here we report on the limitations of K14/K5 as a limbal stem cell marker. K14/K5 expression was measured by immunohistochemistry, Western blotting and Real time PCR and compared between bovine epithelial cells in the limbus and central cornea. A functional study was also included to investigate changes in K5/14 expression within cultured limbal epithelial cells undergoing forced differentiation. K14 expression (or its partner K5) was detected in quiescent epithelial cells from both the limbal area and central cornea. K14 was localized predominantly to basal epithelial cells in the limbus and suprabasal epithelial cells in the central cornea. Western blotting revealed K14 expression in both limbus and central cornea (higher levels in the limbus). Similarly, quantitative real time PCR found K5, partner to K14, to be expressed in both the central cornea and limbus. Following forced differentiation in culture the limbal epithelial cells revealed an increase in K5/14 gene/protein expression levels in concert with a predictable rise in a known differentiation marker. K14 and its partner K5 are limited not only to the limbus but also to the central bovine cornea epithelial cells suggesting K14/K5 is not limbal specific in situ. Furthermore K14/K5 expression levels were not lowered (in fact they increased) within a limbal epithelial cell culture undergoing forced differentiation suggesting K14/K5 is an unreliable maker for undifferentiated cells ex vivo.
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影响因子：
2.9
作者：
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通讯作者：
Wolosin, JM
影响因子：
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作者：
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通讯作者：
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