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Cytokeratin 8 Is Expressed in Human Corneoconjunctival Epithelium, Particularly in Limbal Epithelial Cells

机译:细胞角蛋白8在人角膜结膜上皮,特别是在角膜缘上皮细胞中表达

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Purpose.: The purpose of this study was to investigate the expression of cytokeratin (CK) 8 in the corneoconjunctival epithelium. Methods.: In 17 cadaveric corneoscleral discs and 3 other discs, the presence of CK8 alone or CK8, together with CK3, CK15, vimentin, and integrin ?±6, was investigated by using indirect immunohistochemistry on radial cryosections. Four corneoscleral discs stored in organ culture were used for the preparation of tangential sections of the limbus and for the isolation of limbal epithelial cells and their subsequent cultivation. CK8 expression was examined by RT-PCR in the corneal, limbal, and conjunctival epithelium. Results.: Sixty percent of the cadaveric corneoscleral samples and all samples stored in organ culture revealed positivity for CK8 in the basal epithelial layer of the limbus. Positive basal cells formed a single line or separated clusters. The signal for CK8 became weaker toward the surface of the limbal epithelium. The colocalization of CK8 with vimentin and CK15 in the limbus was also found. CK3 showed only occasional positivity in some of the surface limbal cells. The expression of integrin ?±6 in the basal membrane was absent or decreased under the CK8-positive clusters. Cell cultures revealed strong positivity for CK8 in approximately 80% of the cultured cells, and CK8 expression in the cornea, limbus, and conjunctiva was determined by RT-PCR. Conclusions.: The study demonstrates the strong expression of CK8 in limbal epithelial basal cells, which is maintained during the differentiation and migration of the limbal cells toward the central corneal epithelium.
机译:目的:本研究的目的是研究角膜结膜上皮细胞角蛋白(CK)8的表达。方法:在放射冷冻切片上采用间接免疫组织化学方法,在17具尸体角膜巩膜片和其他3枚椎间盘中,对CK8或CK8以及CK3,CK15,波形蛋白和整联蛋白α±6的存在进行了研究。储存在器官培养物中的四个角膜巩膜盘用于制备角膜缘的切向切片,分离角膜缘上皮细胞并随后进行培养。通过RT-PCR检查CK8在角膜,角膜缘和结膜上皮中的表达。结果:60%的尸体角膜巩膜样品和所有保存在器官培养物中的样品显示角膜缘基底上皮层中CK8呈阳性。阳性基底细胞形成单线或分离的簇。朝向角膜缘上皮表面的CK8信号变弱。还发现角膜缘中CK8与波形蛋白和CK15共定位。 CK3在某些表面角膜缘细胞中仅偶发阳性。在CK8阳性簇下,基膜中整合素β±6的表达不存在或降低。细胞培养显示出约80%的培养细胞对CK8具有强阳性,并且通过RT-PCR测定了CK8在角膜,角膜缘和结膜中的表达。结论:该研究表明CK8在角膜缘上皮基底细胞中的强表达,在角膜缘细胞向角膜上皮的分化和迁移过程中得以维持。

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